Preface

Cheese is one of the oldest fermented and preserved foods in the world. More than 1500 cheese varieties have been classified to date.

Cheese has great nutritional value, which may be attributed to the elaboration of several bioactive peptides during its ripening. Moreover, cheese contains beneficial fatty acids such as conjugated linoleic. Therefore, cheese is considered a functional food. Recent trends to develop functional cheese varieties are ongoing. Several studies have reported the potential for using the cheese matrix as an excellent delivery vehicle for bioactive peptides, vitamins, minerals, and other innovative functional ingredients. Cheese varieties can respond quickly to consumer's evolving needs for functional foods, creating some exciting opportunities for cheese manufacturers. Some novelty versions of functional cheese have been recently launched, including snacks to provide satiety and support fitness and weight; snacks containing billions of live and active probiotics; cheese treats fortified by vitamins A, B, and D, as well as with prebiotics and postbiotics. Other cheese snacks are fortified with micronized iron, zinc, selenium, PUFA, and polyphenols.

This book provides insight into cheese ripening and its impact on the health benefits of cheese. It also discusses cheese's bioactive peptides and fatty acids, the hidden functional benefits of cheese, and recent trends in camel milk cheese and Indian cheese. This volume is a useful resource for food scientists, food chemists, researchers in human nutrition, functional food and/or cheese manufacturers, and a cheese lovers worldwide.

> **Adham M. Abdou** Department of Food Hygiene and Control, Faculty of Veterinary Medicine, Benha University, Moshtohor, Egypt

Section 1 Introduction

#### **Chapter 1**

### Introductory Chapter: Cheese as a Natural Functional Food

*Adham M. Abdou*

#### **1. Introduction**

Cheese is one of the oldest fermented and preserved foods. It was found in early civilizations. The world's oldest cheese was recently found in an Egyptian tomb, in 2018 researchers published proteomic analyses of the solid white mass as the ancient cheese.

Today, more than 1500 varieties of cheese are classified by milk source, coagulation method, moisture content, texture, ripening or aging, and probiotic content. Cheese has great nutritional value with a good source of proteins, fats, vitamins, and minerals. The health benefits of cheese can be attributed to the formation of several bioactive peptides during its ripening or aging. In addition, cheeses contain healthy fatty acids, such as conjugated linoleic acid and phytanic acid. Therefore, cheese can be considered an optimally functional food. According to the EU, functional foods are foods that contain biologically active components that can improve human health or reduce the risk of disease. Recent trends to develop functional and healthier cheese varieties continue. In addition, cheese would be an ideal choice of functional foods. Several studies have reported the possibility of using the cheese matrix as a carrier for bioactive peptides, vitamins, minerals, and other innovative functional ingredients. In addition, the cheese matrix can act as a potential environment for probiotics because its matrix has a high buffering capacity and a dense protein network that protects probiotics from the harsh stomach environment. The cheese category can quickly respond to the changing needs of consumers for healthy food. This would create exciting opportunities for cheese producers. Recently, some innovations have been introduced to the market regarding cheeses as health food snacks, treats, and other forms. Cheese snacks that support satiety, fitness, and weight; snacks containing billions of live and active LGG probiotics; cheese treats enriched with vitamins A, B, and D; cheese forms supplemented with prebiotic oligosaccharides; and other cheese snacks with added liposomal micronized iron and zinc, selenium, and polyphenol (green tea catechins).

#### **2. Cheese as a functional food with great health benefits**

Cheese is a crucial food for bone health, dental and oral health, and weight management. It contains essential nutrients such as calcium, protein, magnesium, zinc, and vitamins A, D, and K, which are essential for maintaining healthy bones. Cheese proteins and their derived peptides also positively enhance bone health by regulating cellular markers and signaling of osteoblasts and osteoclasts. Cheeseeating can protect against cavities by increasing saliva production during chewing

and preventing erosion. Cheese has the highest anticariogenic properties among dairy products studied. Cheese consumption can also help maintain body weight due to its fat, protein, and various vitamins and minerals. Consuming full-fat cheese was linked with a lower risk of obesity compared to low-fat cheese. Balancing cheese intake with low-energy-dense foods, such as fruits and vegetables, is wise. Cheese can also promote satiety as it enhances the feeling of inhibition over further eating and between meals. High protein content in cheese enhances satiety regardless of fat content, providing the potential for decreased energy intake when included as part of a diet.

Low-sodium cheeses, such as Swiss, mozzarella, ricotta, feta, cottage, parmesan, goat cheese, and low-fat cream cheese, can help lower blood pressure. Cheese proteins are rich in ACE inhibitory peptides that can significantly lower blood pressure. Some research has found a link between a diet containing cheese and lower blood pressure, with studies showing that adding certain cheeses to a diet may lead to a decrease in blood pressure.

Cheese and other dairy products are rich sources of antioxidants, which are essential for brain health and neurodegeneration. Cheese's antioxidant properties are attributed to the degradation of casein during ripening, and probiotic cultures can increase its antioxidant activity. Consuming cheese may protect against sodium-rich foods as it contains proteins and peptides that protect blood vessels from short-term adverse effects.

Cheese found to promote gut health by containing probiotic bacteria, which are necessary for maintaining cholesterol levels. Studies have shown that fermented dairy products can change the intestinal microbiota in favor of the host, promoting the growth of *Lactobacillus* and *Bifidobacterium* bacteria. Additionally, cheese may contain natural bioactive peptides that regulate the gut microbiota.

Cheese can help to prevent common cancers, such as colon and bladder cancer. The calcium content of cheese can help prevent these cancers, and cheese may contain specific anticancer peptides produced during cheese processing. Milk proteins are promising candidates for developing anticancer drugs, and peptides derived from dairy products can be detected in fermented milk products, such as cheese.

Immunity support is another benefit of cheese as it contains conjugated linoleic acid (CLA), which may help reduce inflammation and protect against coronary heart disease and obesity. Full-fat dairy products can be healthy when eaten in moderation, and cheese enriched with probiotic microorganisms can strengthen the immune system and prevent immune aging.

Muscle health is also improved by cheese consumption as it increases muscle protein synthesis. Ingestion of cheese protein leads to increased plasma amino acid concentration and subsequently increases muscle protein synthesis.

#### **3. Cheese as a vehicle for functional ingredients (cheese fortification)**

Cheese fortification increases the number of essential micronutrients in food while improving its nutritional value and providing health benefits with little to no risk. Fortification is the addition of a nutrient, known as a fortifier or additive, to a cheese variety. This nutrient is either absent or present in low concentrations and acts as a carrier within the meal. Fortification aims to eliminate nutrient intake deficiencies and the resulting deficiencies. It aims to achieve a balanced overall nutrient profile, compensate for nutrient losses during processing, and meet the needs of consumers looking to supplement their diet. As a result, cheese fortification is used as a public health practice to increase the consumption of essential nutrients.

#### **3.1 Minerals fortification**

#### *3.1.1 Iron*

A crucial component of human nutrition is iron (Fe). Iron insufficiency is a prevalent and pervasive issue in both industrialized and developing nations. The addition of iron derivatives to food, such as cheese, can be considered a potential strategy for the prevention of iron insufficiency. The nutritional content of dairy products can be enhanced through the fortification of iron (Fe). However, there is a potential for a negative impact on the appeal of cheese. The organoleptic alterations and bioavailability of iron are influenced by various factors, including the physical qualities of the added iron (such as valence, solubility, and degree of chelation), as well as the physical features of iron after processing or storage. The incorporation of iron in Fe-fortified cheese is achieved through the formation of iron-polyphosphate/ whey-protein complexes, ferric casein complex, and the addition of ferric chloride (FeCl3). The process of fortifying cheeses with iron enhances their role as primary sources of dietary iron, hence decreasing the prevalence of iron insufficiency. If all cheeses were fortified, there would be a 14% increase in the average intake of Fe. Research conducted on Cheddar cheese indicates that it is feasible to fortify the cheese with iron without adversely affecting its quality; cheddar cheese remains unaffected by any iron supply, even after a 12-month aging period. Recently, a micronized, dispersible ferric pyrophosphate (Sunactive Fe) has been used in Japan to fortify milk and milk products. Milk products have previously been shown to be difficult to fortify with readily absorbable iron due to organoleptic problems. Published studies showed that iron absorption from Sunactive Fe is similar to that of ferrous sulfate from a fortified infant cereal, as well as from fortified dairy products. The high relative bioavailability is presumable due to the very small particle size. Micronized dispersible ferric pyrophosphate can be expected to provoke fewer unacceptable sensory changes than water-soluble iron compounds in different food vehicles; therefore, some cheese varieties fortified with Sunactive Fe have been launched in the Japanese market.

#### *3.1.2 Zinc*

Zinc is a vital micronutrient that is naturally occurring in various food sources. The human body cannot store zinc and hence necessitates regular consumption of this mineral to sustain overall well-being and preserve optimal health. Foods that are consumed in significant quantities, such as cheese, are regarded as preferred vehicles for fortifying food products with low zinc concentrations. Hence, the fortification of cheese with zinc presents a commendable opportunity to enhance the nutritional value of the diet for individuals who are susceptible to certain health risks. Zinc supplementation has been found to offer a modest level of antioxidant protection throughout the maturing process of Cheddar and Edam cheeses. Additionally, it has been shown that the inclusion of zinc-fortified Edam cheese has the potential to enhance the maturation process and enhance the sensory characteristics of the product. In Japan, there has been a recent utilization of a micronized and dispersible form of Zinc oxide known as Sunactive Zn to fortify various food products, including select varieties of cheeses. SunActive Zn fortified cheese offers enhanced bioavailability, prevention of precipitation, increased stability, and elimination of any undesirable taste associated with zinc.

#### *3.1.3 Selenium*

Milk proteins can effectively retain selenium, resulting in its subsequent transfer during the cheese-making process. Selenium-containing amino acids, such as Se-methionine, can be readily integrated into milk proteins, so rendering cheese a favorable dietary source of selenium. Caciocavallo cheese, which is produced using milk that has been enriched with Se. This particular cheese variant exhibits an increased concentration of linoleic acid and conjugated linoleic acid (CLA). Also, it has been reported that fortification of Turkish white cheese with Se through fortification of brine solution during the ripening process, yielded the highest selenium recovery, this resulted in a recovery rate of 70.91%.

#### **3.2 Vitamins fortification**

Vitamin D is frequently utilized as a fortifying agent in cheese, whereas vitamins A, C, and E are commonly found in other sectors of the food industry. The compounds C and E have been found to possess properties that stabilize fat and may potentially be utilized in the production of cheese. Ricotta cheese, a wellknown and widely consumed Italian fresh whey cheese, is traditionally made from cow's milk. The implementation of food fortification has proven to be an effective approach in mitigating the widespread occurrence of vitamin D insufficiency on a global scale. A published study provides evidence that ricotta cheese serves as a suitable substitute dairy medium for the fortification of vitamin D3. The incorporation of emulsified flaxseed oil enables the fortification of vitamin D3 in cheese. The addition of vitamin D3 and PUFA to cheese resulted in favorable effects on the composition, yield, and chemical stability of the final product. Obtaining vitamin D from dietary sources poses challenges due to its limited natural occurrence in food. The cheese matrix is an effective medium for delivering vitamin D. Hence, the utilization of microencapsulation is essential to maintain the functionality of vitamin D. This technique has significant advantages such as enhanced stability against mechanical and photochemical stress, higher oral bioavailability, and enhanced organoleptic qualities. The thermal stability of the fat-soluble vitamins A, D, and E is noteworthy as they do not undergo degradation with the storage time. The addition of vitamins A and E has been found to offer enhanced protection against lipid peroxidation throughout the three-month aging process of Cheddar cheese, and the incorporation of lecithin has been found to result in a 15% increase in the retention of vitamin A and a 26% increase in the retention of vitamin E. Cottage cheese, which has been enriched with vitamins C and A, has a retention rate of 70% and 75%, respectively, over 2 weeks of refrigeration at a temperature of 3°C. Notably, this retention occurs without any discernible alterations to the cheese's flavor, aroma, or appearance. To develop a fortified cheese with vitamin B12, the use of encapsulation vitamin B12 within double emulsions demonstrated an efficiency exceeding 96% and effectively prevented the loss of vitamin content during simulated stomach digestion in laboratory settings. The double emulsion stabilized with sodium caseinate exhibited a release efficiency of less than 5% for encapsulated vitamin B12. Specifically, the encapsulation process boosted the retention rate from 6.3% to over 90% in fortified cheese.

#### **3.3 Addition of bioactive materials**

#### *3.3.1 Omega 3 and polyunsaturated fatty acids (PUFAs)*

The incorporation of polyunsaturated fatty acids (PUFAs) into cheese occurs during the cheese-making process. The oxidation of these highly unsaturated fatty acids leads to the production of distinct odors such as oxidized, rancid, or fishy odors. Microencapsulation is employed as a technique to conceal undesirable sensory attributes and safeguard polyunsaturated fatty acids throughout their processing. The use of flaxseed oil, which is abundant in alpha-linolenic acid, enables the enhancement of cheese with omega-3 and other polyunsaturated fatty acids (PUFAs). The utilization of calcium caseinate for stabilizing flaxseed oil particles has resulted in enhanced resistance against lipid peroxidation and improved chemical stability in fortified cheese. The addition of omega 3 and PUFAs to cheese resulted in favorable effects on the composition, yield, and chemical stability of the resultant fortified cheese. A recent study reported successful trials to fortify food with nanoliposomal encapsulated omega-3 and PUFAs, and another study showed the increase of the bioavailability of omega-3 in fortified dairy products by nanoemulsion of algal oil rich in omega 3.

#### *3.3.2 Conjugated linoleic acid (CLA)*

Conjugated linoleic acid (CLA) refers to a group of isomers that are found in meat and dairy products obtained from ruminant animals. The cis-9/trans-11 and trans-10/cis-12 isomers of CLA are considered to be bioactive. Several investigations have identified alternative CLA isomers in aged cheeses. In the field of fortification, the incorporation of CLA poses a challenge due to its hydrophobic nature. Hence, in aquatic environments, it is more advantageous to introduce the product in the form of an emulsion. Moreover, it is imperative to homogenize the fortified milk before the cheese production process to provide a uniform dispersion and stabilization of fatty acids.

#### *3.3.3 Gamma amino butyric acid (GABA)*

GABA is an amino acid that works as a neurotransmitter inhibitor inside the central nervous system of mammals. GABA has been demonstrated to have significant impacts on brain function, including the potential to mitigate or prevent conditions such as anxiety, depression, insomnia, and memory impairment. Additionally, GABA has been found to boost the immune system, inhibit inflammation processes, and potentially offer protective effects against hypertension and diabetes. GABA is found naturally in trace amounts in several plant-based meals and is particularly abundant in fermented items, such as fermented dairy products, such as cheese. Several studies have documented the capacity of specific strains of lactic acid bacteria (LAB) and bifidobacteria to synthesize GABA. It has been reported the capacity of *Lactobacillus brevis* BT66 (referred to as DSM 32386) and *Streptococcus thermophilus* 84C to generate significant levels of GABA. These strains can create GABA in cheese during the ripening process. Recently, a natural GABA has been produced by natural fermentation in Japan and added at a standardized dose to enrich some kinds of functional

foods, including some varieties of cheese. The addition of natural GABA allows customers to get the health benefits of GABA in their functional cheese.

#### *3.3.4 Coenzyme Q10 and ubiquinol*

Coenzyme Q10 (CoQ10) is a potent natural antioxidant that plays a crucial function in cellular bioenergetics and has been associated with a wide range of established health advantages. The fortification of food products has the potential to substantially boost intake. However, achieving this goal has been challenging, especially with low-fat, water-based products, mostly due to their lipophilicity. Various forms of coenzyme Q10 (CoQ10) with enhanced water solubility or dispersibility have been formulated to enable the enrichment of aqueous products and enhance their bioavailability. The bioactivity of CoQ10 can be maintained when it is integrated into cheese. The encapsulation of coenzyme Q10 in a simple emulsion could improve the stability of CoQ10 in cheese.

Ubiquinol, a bioactive form of Q10, has enhanced absorption properties and is commonly utilized in the production of dietary supplements. In Japan, ubiquinol serves as an effective fortifying agent for functional foods, including several forms of functional cheese.

#### *3.3.5 Collagen peptides*

Collagen peptides, found in dairy products such as cheese, have various applications in addressing skin aging, managing conditions, such as osteoarthritis, and improving nail strength. They are particularly useful in cheese production, where hydrolyzed collagen can enhance structural, mechanical, and health-related characteristics, and play a crucial role in the management and prevention of osteoporosis and related ailments.

#### *3.3.6 Carotenoids*

Lutein-fortified cheeses provide daily carotenoid intake and can help prevent oxidative stress-related illnesses. Lutein-enriched cheeses can preserve physicalchemical, microbiological, and sensory attributes without significant changes. Lutein exhibits favorable stability characteristics during storage, rendering cheeses a suitable carrier for delivery.

#### *3.3.7 Polyphenols*

Polyphenols, such as green tea polyphenols, have antioxidant properties and can interact with protein, carbohydrates, and lipids, affecting the nutritional composition of cheese. The cheese curd matrix exhibits a significant capacity for the retention and preservation of polyphenols that are incorporated into it.

#### *3.3.8 Other additives*

Various additives as plant and animal by-products have been reported in many published researches. The fortification aimed to enhance cheese organoleptic, sensory, and health benefits of the fortified cheeses. Ingredients, such as tomato, cranberry, green tea, broccoli, grape, Morinaga, asparagus, saffron, hibiscus, and others, have been published.

#### **3.4 Probiotics fortification**

Probiotics are live microorganisms that can improve the health of a host when administered in adequate quantities. Cheese is an ideal carrier for probiotics due to its protective barrier against acidic conditions in the gastrointestinal tract (GIT) and its high lipid content, which provides additional safeguarding during transportation. Probiotics can be used in cheese fortification with other strategies, such as iron or zinc fortification, to enhance sensory attributes and make "multi-fortified" cheese a commercially viable product. Cottage cheeses offer several advantages over other dietary options, including pH, fat content, mechanical consistency, low oxygen level, less demanding technological requirements, non-matured state, cold storage, and short shelf-life. Probiotics compete with pathogenic bacteria within the gastrointestinal tract, producing peptides called bacteriocins with antibiotic-like properties. These enzymes inhibit the growth of pathogenic bacteria, such as *Listeria monocytogenes*. Probiotics can incorporate enzymes into the host organism, exhibiting metabolic activities such as protease, lipase, esterase, amylase, and other enzymatic functions. They also facilitate the enzymatic conversion of indigestible carbohydrates into short-chain fatty acids, which have therapeutic properties for gastrointestinal disorders and decrease the pH of the intestinal environment. Probiotics synthesize vital vitamins, bioactives, and antioxidant enzymes and have favorable characteristics in cheese processing and preservation. The selection of strains plays a crucial role in probiotic development. Techniques used in cheese manufacturing significantly impact the viability of probiotic microorganisms, including inoculation methods, flavorings, competition and antimicrobial presence, pH and temperature conditions, preservation methods, salt, and packing materials.

#### **3.5 Prebiotics fortification**

Prebiotics are nondigestible food components that give probiotic microorganisms nourishment, increasing their chances of survival and implantation in the host digestive system. Inulin and fructooligosaccharides (FOS) are significant prebiotic components in foods, providing nourishment to probiotic microorganisms and enhancing their survival and implantation in the host's digestive system. Inulin and FOS have been used in cheese making to create reduced-fat cheese additives with prebiotic properties and dietary fiber. Incorporating inulin into soft cheese and cream cheese yielded lower fat content and elevated moisture levels, with a satisfactory resemblance to the control cheeses. However, the retention of inulin was inadequate to achieve the necessary functional properties. Galactomannans, hydrophilic polysaccharides obtained from legume seeds, have nontoxic characteristics and emulsifying and gelling qualities, facilitating the creation of a protective coating. Chitosan, a biopolymer derived from crustacean exoskeletons, has inhibitory properties against bacteria and fungi proliferation and can be enhanced by fatty acids. Its desirable attributes include biodegradability, biocompatibility with human tissues, and non-toxicity. Incorporating FOS in reduced-fat formulations indicates a potential resemblance to the structure and overall qualities of the original full-fat cheese. However, achieving an exact replication of full-fat cheese after fat reduction is challenging, and the inclusion of FOS presents a technological obstacle.

#### **4. Cheese novelties as healthy snaking trends**

As a result of its natural nutritional advantages, such as its high protein and calcium content, the recent trend to develop functional and/or functional cheese, therefore, cheese is becoming more and more popular as a snack food. Cheese is a product that offers opportunities for creativity in terms of taste, format, texture, and flavor, while also being considered luxurious. Cheese has the potential to rival other widely consumed protein snacks, such as nutrition bars and jerky, in terms of its ability to promote satiety and contribute to fitness and weight management goals as a nutritious protein snack, particularly when consumed in a single-serve portion.

The impetus behind advancements in cheese production is driven by the growing inclination toward nutritious snacking, a phenomenon that is propelled by the demands of fast-paced lifestyles and the increasing urbanization of society. In the context of smaller sizes, cheese producers prioritize the examination and promotion of the nutritional advantages associated with their products.

Cheese snacks are a suitable choice for health-conscious individuals seeking nutritious snacking alternatives as they offer a natural source of high protein and low-fat milk along with additional health advantages. A recently emerged category within the cheese snack market is that of solid shelf-stable cheese snacks (SSSCs). These snacks are comprised solely of dehydrated natural cheese, rendering them both nourishing and satisfying, while also possessing a convenient and enticing texture. To enhance their attractiveness, SSSCs are available in a variety of dimensions, configurations, arrangements, and tastes. The user did not provide any text to rewrite. Nevertheless, it is imperative to conduct additional studies to enhance nutrition and intestinal health, decrease sodium levels, and advocate for sustainable manufacturing methods. The exploration of innovations in novel packaging materials is equally deserving of attention.

The opportunities to innovate with cheese snacks are infinite, as cheese pairs well with salty, savory, and sweet accompaniments. Cheese also may be cut and formed into many shapes and sizes. Moreover, cheese can be loaded with nutrients, namely protein and calcium, which appeal to health- and wellness-conscious consumers.

Cheese snack innovations are on the rise. Cheese may be cut and formed into many shapes and sizes healthy and functional cheese varieties would be available in the form of small portions, bites, balls, slices, sachets, and others to offer a dose-controlled fortified and/or functional cheese, which appeals to health- and wellness-conscious consumers.

#### **5. Cheese as a rich source of valuable materials**

#### **5.1 Calcium**

Cheese is a great dietary source of calcium, essential for sustaining life and regulating vascular function, neuronal transmission, muscle function, and hormone secretion. Only 1% of the total calcium is needed for specific physiological processes, while the remaining 99% is primarily sequestered within the skeletal system. Calcium intake is around 1000 mg for the average adult.

#### **5.2 Proteins**

Cheese is also a great source of protein, essential for the body's formation, regulation, repair, and protection. Approximately 1–2 servings of protein-rich foods per day are sufficient for most adults. Parmesan cheese is the most protein-rich option, with one ounce providing seven grams.

#### **5.3 Vitamin K2 and B12**

Cheese is rich in essential vitamins K2, B12, and B12, which are crucial for hemostasis and brain function. Vitamin K2 is less emphasized than K1, but it interacts with calcium and vitamin D. Hard cheeses, such as gouda and brie, have higher levels of vitamin K2, with gouda and brie having the highest levels. Vitamin B12, the largest and most complex vitamin, is essential for erythrocyte synthesis, protein synthesis, and cognitive processes. Cheeses, such as Swiss cheese, have the highest concentration of B-12, providing 39% of the recommended daily intake.

#### **5.4 Healthy fats**

Cheese, in moderation, can help you get these necessary fats into your diet. Try choosing aged cheeses, such as parmesan, and using it as a garnish for salads. The fats in the cheese will help keep you full and help your body absorb the vitamins in your vegetables.

#### **5.5 Conjugated linoleic acid (CLA)**

Conjugated linoleic acid (CLA) is a complex molecule often underappreciated due to low-fat and no-fat dietary patterns. CLA is a vital component of a healthy diet, often found in dairy and meat products derived from grass-fed ruminant animals. It aids in reducing adipose tissue, promoting lean muscle mass, and supporting immune and inflammatory systems. Cheese derived from grass-fed cows typically contains high levels of CLA, which is positively correlated with fresh grass consumption.

#### **5.6 Gamma amino butyric acid (GABA)**

GABA would occur naturally in various types of cheese as a byproduct of certain starter cultures. The concentration of GABA found in 22 varieties of Italian cheese ranged from 0.260 to 391 mg/kg, and from 320 to 6773.5 mg/kg in Cheddar cheese. The GABA concentration detected in the latter studies ranged between 15 and 5000 mg/kg, even though L-glutamate was added to milk before starting the fermentation process. Also, it has been reported that *Lactobacillus brevis* and *Streptococcus thermophilus* isolated from traditional alpine cheeses were capable of producing high concentrations of GABA.

### **Author details**

Adham M. Abdou Department of Food Hygiene and Control, Faculty of Veterinary Medicine, Benha University, Moshtohor, Egypt

\*Address all correspondence to: dradham@yahoo.com

© 2024 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### Section 2

## Cheese Ripening and Bioactive Peptides

#### **Chapter 2**

## Cheese Ripening: Impact on Cheese as a Functional Food

*Dina A.B. Awad and Adham M. Abdou*

#### **Abstract**

One of the most popular types of fermented dairy products is cheese. The process of cheese aging is essential for improving cheese quality, and health benefits. Ripened cheese at different times acquired wide diversity of characteristic aromas and textures due to establishing a cascade of intrinsic complex biochemical and metabolic outcomes, resulting in a dynamic shift in microbial flora. Various functional bioactive compounds could be released during the cheese ripening process. Many strategy approaches are employed to accelerate cheese ripening based on increasing lipolysis and proteolysis rate. During cheese aging, microbial spoilage as early and late blowing may occur so, designing smart ripening rooms are very essential equipped with computerized monitoring systems including sensors, software platforms, temperature, and humidity data loggers.

**Keywords:** cheese ripening, cheese peptides, factors affecting cheese ripening, cheese microbiota, bioactive ingredients in cheese

#### **1. Introduction**

Cheesemaking depends on the concentration of milk proteins with or without milk fat. After the salting and packing processes, cheese acquired a longer shelf life. Around the world, cheeses can be produced using a variety of milk sources, processing methods, starter culture, coagulants, and ripening conditions, giving rise to a large number of variants with a vast diversity in terms of texture, flavor, and shape [1]. Cheese ripening (aging or maturing) is the most crucial industrial stage in cheese technology, which establishes a cascade of complex biochemical steps, producing a wide variety of microbial flora and different volatile compounds. This is the critical stage during which the cheese's firmness, aroma, flavor, and other specific cheese characteristics are acquired. Ripening occurs under temperature and humidity circumstances that differ depending on the type of cheese. The longer the cheese ripens, the less moisture it retains and the firmer and stronger-tasting it becomes. Cheese kinds are actually defined based on how they ripen; Brie, Camembert, and Roqueforti are examples of mold-ripened cheese, while Limburger and Tilsit are examples of surface-ripened cheese. Internally-ripened cheeses fall into six categories: semi-hard cheeses like Monterey Jack, hard cheeses like Cheddar and extra-hard cheeses like Parmesan and Asiago; pasta filata, which includes mozzarella and provolone; highsalt cheeses like Feta; cheeses with eyes like Dutch types (Gouda and Edam) and Swiss types (Emmental and Gruyere) [2]. The curd's remaining citrate is metabolized by some citrate-positive lactic acid bacteria (LAB), like *Lactococcus lactis subsp. Cremoris,* and *Lactococcus lactis subsp. lactis biovar. diacetylactis,* producing a variety of taste compounds, including acetoin, acetate, diacetyl, 2-butanone, and 2,3-butanediol, which are linked to the development of Dutch cheeses [3].

The development of flavor and sensorial characteristics in cheese is significantly influenced by ripening conditions, especially the time factor. Each variety of cheese has a distinct volatile chemical at a variable concentration, and flavor which could be measured by a diverse series of methodologies, and computational and descriptive approaches [4, 5]. Several bioactives and healthy peptides are produced from milk components during ripening as a result, mainly, of its degradation by starter cultures endo and exo-enzymes [6].

#### **2. Factors affecting the cheese ripening process**

#### **2.1 Cheese microbiota**

In the cheese industry, characterization of the cheese microbiota is crucial, since some specific microorganisms improve cheese characteristics while others may decrease quality [7]. The cheese microbiota is a sophisticated ecosystem that can develop from raw milk, acidifying starters, and adjunct cultures, as well as a group of prokaryotic, eukaryotic, and viral populations that may arise from machinery and the environment of the cheese manufacturing plant.

LAB are the majority microbiota with a significant role during cheese production and ripening. The shape, numbers, and proportions of LAB are then shaped by cheese making and ripening which exert a selection pressure on microorganisms. Diverse microbiota members may interact cooperatively and competitively, which could have an impact on ripened cheese's rheological, organoleptic, and safety properties [8, 9]. The progression of various microbial species and their interaction during the cheesemaking and -ripening processes, in particular, are crucial for the formation of the distinctive sensory characteristics of each cheese variety [10, 11].

Notably, LAB are a kind of Gram-positive bacteria with the ability to withstand acidic pH, and they are primarily cocci or rods at the microscopical examination. The LAB can convert milk carbohydrates to lactic acid. There are more than sixty genera of LAB, the most common genera found in food for fermentation are *Lactobacillus*, *Leuconostoc, Pediococcus, Enterococcus, Lactococcus, Streptococcus*, and *Weissella* [12, 13]. The natural bacterial population of LAB in milk is the fundamental basis for the manufacture and technology of raw milk cheeses. A recent study conducted in Italy indicates that the microbial population of milk, particularly that used to produce Grana Padano PDO cheese, may be impacted by the multifaceted nature of farming practices. The implementation of stringent hygiene measures and management levels in the dairy farm affects negatively the presence of diverse milk microbiota and decreases bacterial load [14].

LAB play a significant role in raw milk cheeses like Parmigiano Reggiano (the long-ripened hard cheeses) both as starter cultures (SLAB) during curd acidification and as non-starter cultures (NSLAB) during cheese-ripening [15–17].

The majority of facultative heterofermentative lactobacilli, or other terms called NSLABs, are frequently isolated from cheese [18]. For instance, one common NSLAB isolated from long-ripened, hard-cooked cheeses is the *Lactobacillus casei*

group, which includes *Lacticaseibacillus rhamnosus*, *Lacticaseibacillus paracasei*, and *Lacticaseibacillus casei* species. These species contribute to the formulation of the distinctive cheese flavor during the ripening process [19].

During cheese ripening, a complicated sequence of activities happens, resulting in a series of biological reactions. Proteolysis is one of the most critical processes, which is launched by the starter, followed by non-starters, and completed by proteolytic enzymes secreted by the bacterial population. These mechanisms cause conformational changes in the particular peptides and amino acid composition, which continually alters during the aging process. The author Sforza et al. [20] discovered a high association between peptide evolution and enzyme activity, allowing for the classification of cheeses based on their ripening circumstances.

Another earlier study established a relation between the hard cheese Parmigiano Reggiano microbiota with the proteolysis rate that takes place during ripening, which is the cause for the valuable and high-quality characteristics of long-ripened cheeses. The study found the raw milk microbiota for different samples from various dairies were quite similar on the curd level. Once ripening time progressed the microbial composition changed, revealing significant differences between cheese samples at early ripening storage time (1 and 2 months) and at late ripening storage (time 7 and 9 months old). These were attributed to NSLAB species, which are more correlated with different peptide profiles and the significant difference in kinetics and activities of the proteolytic enzymes. This study underlines the critical role of NSLAB in ripened cheese proteolysis. Additionally, the potential use of several peptides as indicators of a specific microbial composition allows for the preservation and valorization of cheese's specificity and connection to its production location [21].

The reaction between propionic acid bacteria and LAB influences the organoleptic properties of the produced final cheese product as texture, flavor, and ripening stability. Specific inoculation doses from *Propionibacterium freudenreichii* strains, one of the propionic acid bacteria spp., usually chosen, are utilized for the manufacture of Emmental cheeses and other Swiss-type cheeses which under propionic acid fermentation produce characteristic taste with distinctive eye formation [22].

A study investigated that the amount of free lysine, glycine, and methionine that is available in cheeses is influenced by propionibacteria and LAB. The highest content of free amino acids was presented in the mature cheeses that were produced by combining mesophilic LAB *Propionibacterium* and *L. casei* strains*.* Compared to the mesophilic LAB starter culture or the addition of *L. casei*, it was clear that the addition of *Propionibacteria* had a greater impact on the free amino acid content in matured cheese samples. Notably, this study demonstrates that the growth of *Propionibacteria* in milk is exaggerated by mesophilic LAB, resulting in the production of a significant amount of free amino acids [23]. Not only changing the cheese's body and texture and forming the flavors that give it its distinct character, chemical reactions that take place on the surface of the cheese during ripening also provide an extra protection layer against dry storage conditions [24]. Furthermore, during maturation, the surface microbiota grows quickly, and the residing microorganisms have inhibitory characteristics that hinder pathogenic foodborne bacteria or molds that produce mycotoxin from colonizing the cheese [25, 26]. Numerous enzymes can initiate chemical reactions which may originate from natural milk enzymes, rennet extract utilized in manufacture, or produced by bacteria that survive pasteurization or are added during the process of manufacture or ripening. For instance, three distinct strains of *Brevibacterium linens* were used as surface inoculants to initiate the formation of red smears during the fermentation of Munster cheese [27]. Environmental

pathogens pose a significant risk to the ripening and manufacturing processes of bloomy rind cheeses, such as Camembert, Brie, and related varieties. Due to the product's exposure to the open air during ripening, this risk is especially elevated [28]. The most growing fungi include *Penicillium candidum*, *Kluyveromyces marxianus*, and *Geotrichum candidum* [29]. All predominantly used fungi have a great role in ripening and characteristic organoleptic properties [30].

Cheeses that have been ripened by mold may be distinguished into two main types: those with blue veins and the other with surface mold-ripened cheeses or bloomy rinds [31]*.* The development of a particular mold called Penicillium roqueforti gives Roqueforti, also referred to as blue-veined cheeses, characterized by unique flavor and appearance. Roqueforti cheese is produced in several cities across the globe. Each of these countries' distinct blue cheese varieties has its own characteristics and methods of manufacture [32].

The microbiota of cheese has been shown to have anti-cancer and cholesterollowering capabilities in addition to participating in the enhancement of cheese flavor through the synthesis of volatile molecules [33–35]. The cheese microbial dynamic is affected by the interactions among some factors as LAB used as SLAB or NSLAB, cheese-making processes, and some storage conditions [36–38]. Owing to the cheese microbiota being correlated to the quality and physicochemical properties of cheese, it became critical to understand the cheese microbial properties. In a study reported by Choi et al. [39], the post-inoculation cheese microbiota was found to be dominated by SLAB, and the authors observed that the addition of SLAB resulted in modifications to the microbial community structure, microbial diversity, biomarkers, and predicted functional qualities. Additionally, 105 and 119 days after age, undefinable *Lactobacillus*, or NSLAB, were found.

#### **2.2 Enzymes**

The primary factor in turning milk into cheese is the presence of enzymes, which can be found in the milk itself or introduced as rennet or microbial enzymes. The disintegration of caseins is by far the most significant of the enzymatic processes. Rennet, native milk proteinase, and peptidases generated by starter cultures, enzymes of secondary starters, and enzymes of non-starter cultures are the five primary systems that aid in the hydrolysis of casein [40].

Depending on the cheese type, the ripening time for cheeses made with rennet might range from a few weeks to several years. Microbiological and biochemical alterations take place during ripening, giving the variety's distinctive flavor and texture their development. Primary (lipolysis, proteolysis, and metabolism of residual lactose, lactate, and citrate) or secondary (metabolism of fatty acids and amino acids) processes can be used to categorize biochemical changes in cheese during ripening. Early in the ripening process, lactate is quickly produced from residual lactose. An essential precursor for several processes, such as racemization, oxidation, and microbial metabolism, is lactate. In certain types, the metabolism of citrate is quite important. Cheese's lipolysis is aided by lipases derived from several sources, especially milk [5].

Cheese ripening can be accelerated by increasing the concentration of important enzymes used in cheese production. Plasmin, chymosin, and intracellular and/or cell wall proteinases and peptidases of the LAB and NSLAB are among the proteinases and peptidases found in cheese. Proteases and lipases from animals or fungi are frequently utilized in the production of enzyme-modified cheese. It is uncommon, though, for

*Cheese Ripening: Impact on Cheese as a Functional Food DOI: http://dx.doi.org/10.5772/intechopen.114059*

these enzymes to be used directly to enhance the flavor and ripening of cheese. The primary disadvantages of this approach are the limited availability of commercial enzymes that have been approved for use in cheese ripening and the incapacity to blend the enzymes into the cheese matrix. Exogenous enzymes can be either single enzymes or commercial enzyme combinations. Exogenous enzymes can be added directly into cheese blocks, in combination with starter cultures or coagulants, with cheese-milk, or at the stage of dry salting. When making cheddar cheese, the latter process is employed [41]. Exogenous enzymes were supposed to accelerate cheese ripening and aid in the production of unique tastes in specific cheese varieties. In that regard, a few examples of distinct enzymes or their mixes were reviewed and documented [5].

#### **2.3 Dairy animal feeding**

It was worth noting that dietary supplementation of lactating dairy cows may change the quality of dairy products, especially cheese products. The characteristic volatile substances that contribute to cheese flavor were attributed to the aromatic qualities of milk obtained from lactating dairy ruminants. Extensive research has been conducted concerning lactating dairy ruminants fed specific experimental diets, such as those distinguished, for example, by the addition of trace elements, natural supplements, or agricultural byproducts rich in bioactive compounds. Cheese contains a variety of volatile substances, such as carboxylic acids, lactones, ketones, alcohols, and aldehydes. The relative amounts of each substance are determined by the biochemical processes that take place during ripening. These processes are primarily mediated by endogenous enzymes and elements of bacterial origin whose function can be greatly influenced by the bioactive substances consumed by animals in their diet and released in milk through the mammary gland. According to Ianni et al. [42] there was a significant correlation between the quality of the biochemical changes in cheese products throughout ripening and the various dairy animal feeding practices.

#### **2.4 Level of sodium chloride**

It is well-recognized that consuming too much sodium raises the risk of hypertension, cardiovascular disease, and even stomach cancer [43, 44]. The issue for the food industry is reformulating food products that contain less sodium aiming to offer low-salt food in the human diet is currently one of the top priorities for public health organizations [45]. To comply with the World Health Organization's (WHO) recommendation of 2 g/day, it has been suggested by the World Health Organization that sodium intake be reduced by 30% [46].

Food businesses must carefully re-evaluate the composition and processing of high-sodium foods considering public awareness of excessive sodium intake and nutrition claims connected to salt content. Although it is usual practice to replace some ingredients in products through reformulation, it is still difficult to reduce salt in cheese due to sodium chloride's various functions and essential activities in cheese making. Salt improves the taste and fragrance profile, controls the texture, final pH, and water activity, and influences microbiological growth. It also favors the drainage of remaining whey. In the end, salt concentration affects the shelf-life of cheese by regulating the activity of starter and non-starter LAB during cheese production and ripening. Any adjustment to the salting process, such as lowering the sodium chloride (NaCl) amount or substituting alternative salting agents, could upset the delicate balance within the parameters, changing the cheese's quality. Depending on the kind

of cheese and manufacturing method (for example, soft, semi-hard, hard, and moldripened cheeses), the decrease of NaCl content may be treated differently. As a result, specific tactics could be implemented to preserve the general quality and safety of various cheese types [47].

As a result, making cheeses with less NaCl content is becoming increasingly popular in the dairy business. Since NaCl is essential for textural qualities, microbial development, autolysis, enzyme activity during ripening, and ultimately cheese flavor, hence, reducing NaCl in cheese-making poses a number of technological, sensory, and microbiological issues [48]. Reduced NaCl semi-hard cheeses typically have enhanced cohesiveness, adhesiveness, acidity, bitterness, and an unpleasant aftertaste along with a decrease in salt and hardness as their sensory defaults [49, 50]. Additionally, reducing the NaCl concentration of cheese affects product safety due to the possibility of germs like *Listeria monocytogenes* growing in the product [51].

According to popular perception, cheese contains varying quantities of salt, depending on the type of cheese. NaCl content for soft, semi-hard, and hard cheeses ranges from 0.5% to 2.5%, whereas it ranges from 3% to 5% for blue-type cheeses. In cheese, NaCl performs a variety of crucial tasks, including modifying the curd and rind's physical characteristics, regulating the microbiota's growth as the cheese ripens, preventing the formation of infections or spoilers, and enhancing the flavor [52, 53].

In a study followed the strategy of reduction of NaCl or its partial substitution with other salts such as potassium chloride in a semihard cheese (Reblochon), it was reported that lowering the salt level in the semi-hard cheese samples caused spoiler growth to accelerate, as seen by increased Pseudomonas species formation and increased cheese proteolysis and lipolysis [45].

#### **3. Bioactive components produced during cheese ripening**

Bioactive peptides are released during the ripening process of cheese and are protein fragments in the form of short amino acid sequences. During cheese maturation, various functional bioactive compounds could be generated such as volatile fatty acids, exopolysaccharides, vitamins, organic acids, peptides, and amino acids (γ-aminobutyric acid and conjugated linolenic acid). Many laboratory and animal research studies demonstrated that most of these generated bioactive compounds exhibited different biological activities including antihypertensive, antioxidant, anticancer, and antimicrobial activities [54–56]. The above bioactivities lead to health-protective effects associated with a reduced incidence of cardiovascular disease risk factors, such as obesity, dyslipidemia, and type 2 diabetes [57], as well as reduced incidence of metabolic syndrome [58, 59]. The artisanal methods employed for the manufacturing of Mexican cheeses, depending on natural milk indigenous microflora, may offer many potential health benefits. A previous study [60] demonstrated that peptides derived from both fresh cheese and a model cheese (laboratory scale) had an antihypertensive peptide through suppressing ACE-inhibitory effect. Although artisanal cheese is under storage conditions, producing different product varieties such as (Crema de Chiapas, Cocido, and Fresco of Sonora). Few limited studies are available for artisanal cheeses in Mexico and the various derived bioactive compounds in artisanal cheeses, and those available focus primarily on the antioxidant and ACE activity of water-soluble extracts (WSE) obtained from different types of artisanal cheese varieties such as (Crema de Chiapas, Cocido, and Fresco of Sonora) from different storage conditions [61, 62].

Distinct peptide sets found in different cheese varieties contribute to their distinctive flavors and possible health benefits. These bioactive molecules support the body's defense against potentially harmful substances by acting as antioxidants, anti-inflammatory agents, and even antimicrobial activities. Angiotensin-converting enzyme (ACE) inhibitory characteristics have been demonstrated by some cheese peptides, indicating that they may lower blood pressure by preventing the angiotensin-converting enzyme from acting [63].

Protein breakdown is typically linked to biological functional qualities and is aided by the action of milk-specific enzymes. Furthermore, the microorganisms used or added during cheese production may produce bioactive peptides [64]. Milk proteases and peptidases produce large and intermediate-sized peptides, which are then hydrolyzed by enzymes from the cheese SLAB and NSLAB strains. These are known as primary proteolysis reactions [65].

Both the body's digestive processes and the fermentation procedures used to produce fermented dairy products produce functional peptides. They are produced by the proteins; casein and whey. When liberated from their original proteins, these peptides take on an active role. When they enter the circulatory system, they may have a systemic effect, or they can be absorbed in intact form and have different physiological effects locally in the gut. It is well known that lactoferroxins and casooxins function as opioid antagonists, while casomorphins and lactophorins generated from milk proteins are opioid agonists. The opioids' analgesic qualities are comparable to aspirin. The actions of casokinins (which lower blood pressure), casoplatelins (which reduce blood clotting), immunopeptides (which boost immune function), and phosphopeptides (which carry minerals) are all examples of casokinin action. Casein phosphopeptides may improve calcium, phosphorus, and magnesium bioavailability for better bone health. They may also aid in the prevention of dental cavities and play a function in the secretion of enterohormones and immunological boosting. The involvement of casein peptides in blood pressure regulation looks promising. Certain casein and whey protein hydrolyzates prevent the conversion of angiotensin I to angiotensin II. Because angiotensin II elevates blood pressure by constricting blood vessels, inhibiting it causes blood pressure to fall. Dairy foods would thus be a natural functional food for managing hypertension due to their ACE inhibitory action. There are several commercially marketed whey products that include discrete bioactive peptides. The glycomacropeptide (GMP) is produced by proteolysis from kappa-casein [5, 63].

Proteins and lipids are the sources of the most common and extensively researched bioactive molecules. Cheese proteins are broken down to form bioactive peptides and amino acids such as gamma-aminobutyric acid and ornithine, while fats are hydrolyzed to produce and be an origin of bioactives like conjugated linoleic acid, carotenoids, fat-soluble vitamins, and sphingolipids, among other things. Depending on their nature, these chemicals exhibit various bioactivities [6, 35].

High quantities of lactic acid and other organic acids are produced by LAB during the fermentation of lactose [66]. The ripening process is a complicated process in which several milk enzymes participate, including rennet and enzymes from LAB, which leads to successive transformations that aim to affect the various curd ingredients [40, 67].

Proteinases found in LAB contribute to the proteolysis of cheese proteins, converting them into oligopeptides that can either be further degraded into shorter peptides and amino acids through the synergistic action of various intracellular peptidases produced by specific LAB. These peptides, amino acids, and their derivatives help the final cheese to develop its texture, flavor, and health benefits. The generated peptides

were subjected to both *in vitro* laboratory experiments and *in vivo* animal models, which confirmed that they possessed a variety of biological activities, including the ability to scavenge free radicals (antioxidants), inhibit microbial growth (antimicrobial), fight inflammation (anti-inflammatory), immunity enhancer (immunomodulatory), opioid blocker receptors activity (analgesic), and lower blood pressure by suppressing the angiotensin-converting enzyme producing antihypertensive effect. Furthermore, some LAB produce vitamins, some antimicrobials, conjugated linoleic acid, and other functional lipids with anti-inflammatory and anticarcinogenic properties, as well as bioactives that contribute to physiological processes like neurotransmission and hypotension induction with diuretic effects [63].

During the early weeks of cheese ripening, NSLAB proliferate at a very slow rate, but eventually take control of the cheese microbiota following the death phase of the starter culture [68, 69]. The process of cheese ripening is sophisticated and a dynamic process. The variety of proteolytic enzymes naturally found in milk and the remaining coagulants, as well as the enzymatic metabolism of LAB, are crucial to this process [70]. Peptides are continuously released during ripening by the action of plasmin and LAB's enzymes; some of these peptides are then digested, while others accumulate throughout storage [71, 72].

One of the major milk groups that can enhance the sensory qualities, shelf life, and microbial safety of cheese as well as the nutritional content and functional features of the finished products is NSLAB. It includes; *Pediococcus*, *Enterococcus*, *Lactobacillus*, and *Leuconostoc* genera. Due to the creation of certain metabolites throughout the ripening process, ripened cheese develops a diversity of flavors, nutritional qualities, and rheological characteristics [73–75]. Peptides, amino acids, biogenic amines, nucleic acids, carbohydrates, organic acids, vitamins, polyphenols, alkaloids, minerals, and


#### **Table 1.**

*List of some published bioactive compounds found in ripened cheese varieties.*

*Cheese Ripening: Impact on Cheese as a Functional Food DOI: http://dx.doi.org/10.5772/intechopen.114059*

any other molecules that can alter the sensory and rheological properties, as well as the nutritional value and health benefits of the finished products, are examples of metabolites [73].

The aforementioned functional biological activities have been linked to a decreased incidence of most common coronary artery disease risk factors, such as dyslipidemia, diabetes type 2, and overweight [57], as well as different metabolic syndromes [59].

As shown in **Table 1** various bioactive components in different ripened cheese.

#### **4. Ripened cheese additives**

One of the simplest and most traditional methods for preserving food to increase its shelf life is the direct addition of additives which may be chemical or naturally derived preservatives. These ingredients are added to cheese in order to prevent spoilage and pathogenic microbial growth, increase shelf life, enhance organoleptic and sensory characteristics, and maintain nutritional value [91, 92].

Food preservatives permitted for use in matured cheeses in the EU are classified into three functional groups: antimicrobials, antioxidants, and antibrowning


#### **Table 2.**

*Additives to cheese milk and its role in cheese ripening.*

chemicals. During cheesemaking, these compounds are added to the milk vat as antimicrobials and antioxidants, or to the cheese as surface defenders against unwanted agents. Lysozyme, sorbic acid/sorbates, nisin, natamycin, hexamethylene tetramine (HTM), nitrates/nitrites, and propionic acid/propionates are authorized additions for matured cheeses. As illustrated in **Table 2** different cheese additives are added to cheese milk for enhancement of the cheese ripening process.

### **Author details**

Dina A.B. Awad and Adham M. Abdou\* Department of Food Hygiene and Control, Faculty of Veterinary Medicine, Benha University, Moshtohor, Egypt

\*Address all correspondence to: dradham@yahoo.com

© 2024 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### **References**

[1] Tekin A, Hayaloglu AA. Understanding the mechanism of ripening biochemistry and flavour development in brine ripened cheeses. International Dairy Journal. 2023;**137**:105508. DOI: 10.1016/J. IDAIRYJ.2022.105508

[2] McSweeney PLH. Cheese: Biochemistry of cheese ripening. In: Wouters JTM, editor. Encyclopedia of Dairy Sciences. 2nd ed. Cambridge, UK: Academic Press; 2011. pp. 667-674. DOI: 10.1016/B978-0-12-374407-4.00080-7

[3] Hassan FAM, El-Gawad MA, Enab A. Flavour compounds in cheese. Research on Precision Instrument and Machinery. 2013;**2**(June):15-29

[4] Ganesan B, Weimer BC. In: McSweeney PLH, Fox PF, Cotter PD, Everett DW, editors. Amino Acid Catabolism and its Relationship to Cheese Flavor Outcomes. San Diego, CA, USA: Academic Press; 2017. pp. 483-516. DOI: 10.1016/ B978-0-12-417012-4.00019-3

[5] Khattab AR, Guirguis HA, Tawfik SM, Farag MA. Cheese ripening: A review on modern technologies towards flavor enhancement, process acceleration and improved quality assessment. Trends in Food Science & Technology. 2019;**88**:343-360. DOI: 10.1016/j. tifs.2019.03.009

[6] Vázquez-García R, Martíndel-Campo ST. Enzyme actions during cheese ripening and production of bioactive compounds. In: Rajput YS, Sharma R, editors. Foundations and Frontiers in Enzymology. CA, USA: Academic Press; 2023. pp. 331-347. DOI: 10.1016/ B978-0-323-96010-6.00012-6

[7] Mayo B, Rachid CTCC, Alegría A, Leite AMO, Peixoto RS, Delgado S. Impact of next generation sequencing techniques in food microbiology. Current Genomics. 2014;**15**(4):293-309. DOI: 10.2174/1389202915666140616233211

[8] Kamimura BA, De Filippis F, Sant'Ana AS, Ercolini D. Large-scale mapping of microbial diversity in artisanal Brazilian cheeses. Food Microbiology. 2019;**80**:40-49. DOI: 10.1016/j.fm.2018.12.014

[9] Mayo B, Rodríguez J, Vázquez L, Flórez AB. Microbial interactions within the cheese ecosystem and their application to improve quality and safety. Foods (Basel, Switzerland). 2021;**10**(602):1-28. DOI: 10.3390/foods10030602

[10] Gobbetti M, Di Cagno R, Calasso M, Neviani E, Fox PF, De Angelis M. Drivers that establish and assembly the lactic acid bacteria biota in cheeses. Trends in Food Science & Technology. 2018;**78**:244-254. DOI: 10.1016/j.tifs.2018.06.010

[11] Afshari R, Pillidge CJ, Dias DA, Osborn AM, Gill H. Cheesomics: The future pathway to understanding cheese flavour and quality. Critical Reviews in Food Science and Nutrition. 2020;**60**(1):33-47. DOI: 10.1080/10408398.2018.1512471

[12] George F, Daniel C, Thomas M, Singer E, Guilbaud A, Tessier FJ, et al. Occurrence and dynamism of lactic acid bacteria in distinct ecological niches: A multifaceted functional health perspective. Frontiers in Microbiology. 2018;**9**:2899. DOI: 10.3389/ fmicb.2018.02899

[13] Mokoena MP. Lactic acid bacteria and their bacteriocins: Classification, biosynthesis and applications against Uropathogens: A mini-review. Molecules (Basel, Switzerland). 2017;**22**(8):1255. DOI: 10.3390/molecules22081255

[14] Bava L, Zucali M, Tamburini A, Morandi S, Brasca M. Effect of different farming practices on lactic acid bacteria content in cow milk. Animals. 2021;**11**:522. DOI: 10.3390/ani11020522

[15] Gatti M, Bottari B, Lazzi C, Neviani E, Mucchetti G. Invited review: Microbial evolution in raw-milk, long-ripened cheeses produced using undefined natural whey starters. Journal of Dairy Science. 2014;**97**:573-591. DOI: 10.3168/jds.2013-7187

[16] Carafa I, Stocco G, Franceschi P, Summer A, Tuohy KM, Bittante G, et al. Evaluation of autochthonous lactic acid bacteria as starter and nonstarter cultures for the production of Traditional Mountain cheese. Food Research International (Ottawa, Ont.). 2019;**15**:209-218. DOI: 10.1016/j. foodres.2018.08.069

[17] Martini S, Conte A, Tagliazucchi D. Effect of ripening and in vitro digestion on the evolution and fate of bioactive peptides in Parmigiano-Reggiano cheese. International Dairy Journal. 2020;**105**:104668. DOI: 10.1016/j. idairyj.2020.104668

[18] Beresford TP, Fitzsimons NA, Brennan NL, Cogan TM. Recent advances in cheese microbiology. International Dairy Journal. 2001;**11**:259-274. Available from: https://www.sciencedirect.com/ science/article/pii/S0958694601000565

[19] Zheng J, Wittouck S, Salvetti E, Franz CMAP, Harris HMB, Mattarelli P, et al. A taxonomic note on the genus lactobacillus: Description of 23 novel genera, emended description of the genus lactobacillus Beijerinck 1901,

and union of Lactobacillaceae and Leuconostocaceae. International Journal of Systematic and Evolutionary Microbiology. 2020;**70**(4):2782-2858. DOI: 10.1099/ijsem.0.004107

[20] Sforza S, Cavatorta V, Lambertini F, Galaverna G, Dossena A, Marchelli R. Cheese peptidomics: A detailed study on the evolution of the oligopeptide fraction in Parmigiano-Reggiano cheese from curd to 24 months of aging. Journal of Dairy Science. 2012;**95**(7):3514-3526. DOI: 10.3168/jds.2011-5046

[21] Bottari B, Levante A, Bancalari E, Sforza S, Bottesini C, Prandi B, et al. The interrelationship between microbiota and peptides during ripening as a driver for Parmigiano Reggiano cheese quality. Frontiers in Microbiology. 2020;**11**:581658. DOI: 10.3389/ fmicb.2020.581658

[22] Fröhlich-Wyder MT, Bisig W, Guggisberg D, Jakob E, Turgay M, Wechsler D. Cheeses with propionic acid fermentation: Chapter 35. In: McSweeney P, Fox P, Cotter P, Everett D, editors. Cheese: Chemistry, Physics, and Microbiology. 4th ed. CA, USA: Academic Press; 2017. pp. 889-910. DOI: 10.1016/ B978-0-12-417012-4.00035-1

[23] Ziarno M, Bryś J, Kowalska E, Cichońska P. Effect of metabolic activity of lactic acid bacteria and propionibacteria on cheese protein digestibility and fatty acid profile. Scientific Reports. 2023;**13**(1):15363. DOI: 10.1038/s41598-023-42633-w

[24] Cogan TM, Hill C. Cheese starter cultures. In: Fox PF, editor. Cheese: Chemistry, Physics and Microbiology. 2nd ed. London: Chapman & Hall; 1993. pp. 193-255. DOI: 10.1007/978-1-4615-2650-6\_6

*Cheese Ripening: Impact on Cheese as a Functional Food DOI: http://dx.doi.org/10.5772/intechopen.114059*

[25] Bockelmann W, Hoppe-Seyler T. The surface flora of bacterial smearripened cheeses from cow's and goat's milk. International Dairy Journal. 2001;**11**:307-314. DOI: 10.1016/ S0958-6946(01)00060-7

[26] Roth E, Schwenninger SM, Eugster-Meier E, Lacroix C. Facultative anaerobic halophilic and alkaliphilic bacteria isolated from a natural smear ecosystem inhibit listeria growth in early ripening stages. International Journal of Food Microbiology. 2011;**147**(1):26-32. DOI: 10.1016/j.ijfoodmicro.2011.02.032

[27] Leuschner RG, Hammes WP. Degradation of histamine and tyramine by Brevibacterium linens during surface ripening of Munster cheese. Journal of Food Protection. 1998;**61**:874-878. DOI: 10.4315/0362-028x-61.7.874

[28] Batty D, Meunier-Goddik L, Waite-Cusic JG. Camembert-type cheese quality and safety implications in relation to the timing of high-pressure processing during aging. Journal of Dairy Science. 2019;**102**(10):8721-8733. DOI: 10.3168/ jds.2018-16236

[29] Sicard M, Perrot N, Leclercq-Perlat MN, Baudrit C, Corrieu G. Toward the integration of expert knowledge and instrumental data to control food processes: Application to camemberttype cheese ripening. Journal of Dairy Science. 2011;**94**:1-13. DOI: 10.3168/ jds.2009-2984

[30] Galli BD, Martin JGP, da Silva PPM, Porto E, Spoto MHF. Sensory quality of camembert-type cheese: Relationship between starter cultures and ripening molds. International Journal of Food Microbiology. 2016;**234**:71-75. DOI: 10.1016/j.ijfoodmicro.2016.06.025

[31] Bates M, Clark S. Mold-ripened cheeses. In: Clark S, Drake M,

Kaylegian K, editors. The Sensory Evaluation of Dairy Products. Cham: Springer; 2023. DOI: 10.1007/ 978-3-031-30019-6\_17

[32] Cantor MD, van den Tempel T, Hansen TK, Ardö Y. Blue cheese. In: McSweeney PLH, Fox PF, Cotter PD, Everett DW, editors. Cheese: Chemistry, Physics and Microbiology. 4th ed. Vol. 2. CA, USA: Academic Press; 2017. pp. 929-954. DOI: 10.1016/ B978-0-12-417012-4.00037-5

[33] Sieber R, Bütikofer U, Egger C, Portmann R, Walther B, Wechsler D. ACE-inhibitory activity and ACEinhibiting peptides in different cheese varieties. 2010;**90**:47-73. DOI: 10.1051/ dst/2009049

[34] Broadbent J, Cai H, Larsen RL, Hughes JE, Welker D, Carvalho VG, et al. Genetic diversity in proteolytic enzymes and amino acid metabolism among *lactobacillus helveticus* strains. Journal of Dairy Science. 2011;**94**:4313-4328. DOI: 10.3168/jds.2010-4068

[35] Potočki S. Potential health benefits of sphingolipids in milk and dairy products. Mljekarstvo. 2016;**66**:251-261

[36] Percival B, Percival F. Reinventing the Wheel: Milk, Microbes, and the Fight for Real Cheese. Berkeley: University of California Press; 2017. DOI: 10.1525/9780520964464

[37] Van Hoorde K, Heyndrickx M, Vandamme P, Huys G. Influence of pasteurization, brining conditions and production environment on the microbiota of artisan gouda-type cheeses. Food Microbiology. 2010;**27**(3):425-433. DOI: 10.1016/j.fm.2009.12.001

[38] Porcellato D, Skeie SB. Bacterial dynamics and functional analysis of microbial metagenomes during ripening of Dutch-type cheese. International Dairy Journal. 2016;**61**:182-188. DOI: 10.1016/j.idairyj.2016.05.005

[39] Choi J, In Lee S, Rackerby B, Frojen R, Goddik L, Ha S-D, et al. Assessment of overall microbial community shift during Cheddar cheese production from raw milk to aging. Applied Microbiology and Biotechnology. 2020;**104**(14):6249-6260. DOI: 10.1007/s00253-020-10651-7

[40] McSweeney PLH. Biochemistry of cheese ripening. International Journal of Dairy Technology. 2004;**57**(2-3):127-144. DOI: 10.1111/j.1471-0307.2004.00147.x

[41] Azarnia S, Robert N, Lee B. Biotechnological methods to accelerate Cheddar cheese ripening. Critical Reviews in Biotechnology. 2006;**26**:121- 143. DOI: 10.1080/07388550600840525

[42] Ianni A, Bennato F, Martino C, Grotta L, Martino G. Volatile flavor compounds in cheese as affected by ruminant diet. Molecules (Basel, Switzerland). 2020;**25**(3):461(1-16). DOI: 10.3390/molecules25030461

[43] Mohan S, Campbell NRC. Salt and high blood pressure. Clinical Science (London, England: 1979). 2009;**117**(1):1- 11. DOI: 10.1042/CS20080207

[44] Song JH, Kim YS, Heo NJ, Lim JH, Yang SY, Chung GE, et al. High salt intake is associated with atrophic gastritis with intestinal metaplasia. Cancer Epidemiology, Biomarkers & Prevention: A Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology. 2017;**26**(7):1133-1138. DOI: 10.1158/1055- 9965.EPI-16-1024

[45] Dugat-Bony E, Bonnarme P, Fraud S, Catellote J, Sarthou A-S, Loux V, et al.

Effect of sodium chloride reduction or partial substitution with potassium chloride on the microbiological, biochemical and sensory characteristics of semi-hard and soft cheeses. Food Research International. 2019;**125**:108643. DOI: 10.1016/j.foodres.2019.108643

[46] WHO (World Health Organization). Guideline: Sodium Intake for Adults and Children. Geneva: Fact Sheet, WHO Publications; 2012

[47] Tidona F, Zago M, Carminati D, Giraffa G. The reduction of salt in different cheese categories: Recent advances and future challenges. Frontiers in Nutrition. 2022;**9**:859694. DOI: 10.3389/fnut.2022.859694

[48] Søndergaard L, Ryssel M, Svendsen C, Høier E, Andersen U, Hammershøj M, et al. Impact of NaCl reduction in Danish semi-hard Samsoe cheeses on proliferation and autolysis of DL-starter cultures. International Journal of Food Microbiology. 2015;**213**:59-70. DOI: 10.1016/j.ijfoodmicro.2015.06.031

[49] Rulikowska A, Kilcawley KN, Doolan IA, Alonso-Gomez M, Nongonierma AB, Hannon JA, et al. The impact of reduced sodium chloride content on Cheddar cheese quality. International Dairy Journal. 2013;**28**(2):45-55. DOI: 10.1016/j. idairyj.2012.08.007

[50] Schroeder CL, Bodyfelt F, Wyatt CJ, McDaniel MR. Reduction of sodium chloride in Cheddar cheese: Effect on sensory, microbiological, and chemical properties. Journal of Dairy Science. 1988;**71**:2010-2020

[51] Hystead EE, Diez-Gonzalez F, Schoenfuss TC. The effect of sodium reduction with and without potassium chloride on the survival of listeria monocytogenes in Cheddar

*Cheese Ripening: Impact on Cheese as a Functional Food DOI: http://dx.doi.org/10.5772/intechopen.114059*

cheese. Journal of Dairy Science. 2013;**96**:6172-6185

[52] Cruz AG, Faria JAF, Pollonio MAR, Bolini HMA, Celeghini RMS, Granato D, et al. Cheeses with reduced sodium content: Effects on functionality, public health benefits and sensory properties. Trends in Food Science & Technology. 2011;**22**(6):276-291. DOI: 10.1016/j. tifs.2011.02.003

[53] Johnson ME, Kapoor R, McMahon DJ, McCoy DR, Narasimmon RG. Reduction of sodium and fat levels in natural and processed cheeses: Scientific and technological aspects. Comprehensive Reviews in Food Science and Food Safety. 2009;**8**:252-268. DOI: 10.1111/ j.1541-4337.2009.00080.x

[54] Faure M, Mettraux C, Moennoz D, Godin J-P, Vuichoud J, Rochat F, et al. Specific amino acids increase mucin synthesis and microbiota in dextran sulfate sodium–treated rats. The Journal of Nutrition. 2006;**136**(6):1558-1564. DOI: 10.1093/jn/136.6.1558

[55] Sprong RC, Schonewille AJ, van der Meer R. Dietary cheese whey protein protects rats against mild dextran sulfate sodium–induced colitis: Role of mucin and microbiota. Journal of Dairy Science. 2010;**93**(4):1364-1371. DOI: 10.3168/ jds.2009-2397

[56] Geurts L, Everard A, Le Ruyet P, Delzenne NM, Cani PD. Ripened dairy products differentially affect hepatic lipid content and adipose tissue oxidative stress markers in obese and type 2 diabetic mice. Journal of Agricultural and Food Chemistry. 2012;**60**(8):2063- 2068. DOI: 10.1021/jf204916x

[57] Sullivan Å, Edlund C, Nord CE. Effect of antimicrobial agents on the ecological balance of human microflora. The Lancet Infectious Diseases.

2001;**1**(2):101-114. DOI: 10.1016/ S1473-3099(01)00066-4

[58] Bonthuis M, Hughes MCB, Ibiebele TI, Green AC, van der Pols JC. Dairy consumption and patterns of mortality of Australian adults. European Journal of Clinical Nutrition. 2010;**64**(6):569-577. DOI: 10.1038/ ejcn.2010.45

[59] Sonestedt E, Wirfält E, Wallström P, Gullberg B, Orho-Melander M, Hedblad B. Dairy products and its association with incidence of cardiovascular disease: The Malmö diet and cancer cohort. European Journal of Epidemiology. 2011;**26**(8):609-618. DOI: 10.1007/s10654-011-9589-y

[60] Torres-Llanez MJ, González-Córdova AF, Hernandez-Mendoza A, Garcia HS, Vallejo-Cordoba B. Angiotensin-converting enzyme inhibitory activity in Mexican Fresco cheese. Journal of Dairy Science. 2011;**94**(8):3794-3800. DOI: 10.3168/ jds.2011-4237

[61] Aguilar-Toalá JE, González-Córdova AF, Hernández-Mendoza A, Torres-Llanez MJ, Vallejo-Cordoba B. Antioxidant activity of bioactive peptides and polyphenols isolated from Mexican Artisanal Cheeses. In: Abstract Number 072-04 in IFT Annual Meeting and Expo, Chicago, IL. Chicago, IL: Institute of Food Technologists; 2013

[62] Santos-Espinosa A, González-Córdova AF, Hernández-Mendoza A, Estrada-Montoya MC, Vallejo-Cordoba B. Angiotensin-converting enzyme inhibitory activity of Mexican Artisanal Cheeses. In: Abstract Number 072-05 in IFT Annual Meeting and Expo, Chicago, IL. Chicago, IL: Institute of Food Technologists; 2013

[63] Santiago-López L, Aguilar-Toalá JE, Hernández-Mendoza A, VallejoCordoba B, Liceaga AM, González-Córdova AF. Invited review: Bioactive compounds produced during cheese ripening and health effects associated with aged cheese consumption. Journal of Dairy Science. 2018;**101**(5):3742-3757. DOI: 10.3168/JDS.2017-13465

[64] Claeys W, Cardoen S, Daube G, Block J, Dewettinck K, Dierick K, et al. Raw or heated cow milk consumption: Review of risks and benefits. Food Control. 2013;**31**:251-262. DOI: 10.1016/j. foodcont.2012.09.035

[65] Chavan RS, Chavan SR, Khedkar CD, Jana AH. UHT milk processing and effect of plasmin activity on shelf life: A review. Comprehensive Reviews in Food Science and Food Safety. 2011;**10**(5):251-268. DOI: 10.1111/j.1541-4337.2011.00157.x

[66] Settanni L, Moschetti G. Nonstarter lactic acid bacteria used to improve cheese quality and provide health benefits. Food Microbiology. 2010;**27**(6):691-697. DOI: 10.1016/j. fm.2010.05.023

[67] Leroy F, De Vuyst L. Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends in Food Science & Technology. 2004;**15**(2):67-78. DOI: 10.1016/j. tifs.2003.09.004

[68] Casey M, Häni J, Gruskovnjak J, Schaeren W, Wechsler D. Characterization of the non-starter lactic acid bacteria (NSLAB) of Gruyère PDO cheese. Lait. 2006;**86**:407-414. DOI: 10.1051/lait:2006020

[69] Zuljan FA, Mortera P, Alarcón SH, Blancato VS, Espariz M, Magni C. Lactic acid bacteria decarboxylation reactions in cheese. International Dairy Journal. 2016;**62**:53-62. DOI: 10.1016/j. idairyj.2016.07.007

[70] Gagnaire V, Daniel M, Herrouin M, Léonil J. Peptides identified during Emmental cheese ripening: Origin and proteolytic systems involved. Journal of Agricultural and Food Chemistry. 2001;**49**(9):4402-4413. DOI: 10.1021/ jf000895z

[71] Ryhänen E-L, Pihlanto-Leppälä A, Pahkala E. A new type of ripened, low-fat cheese with bioactive properties. International Dairy Journal. 2001;**11**(4):441-447. DOI: 10.1016/ S0958-6946(01)00079-6

[72] Ong L, Henriksson A, Shah N. Angiotensin converting enzymeinhibitory activity in Cheddar cheeses made with the addition of probiotic *Lactobacillus casei* sp. 2007;**87**:149-165. DOI: 10.1051/lait:2007004

[73] Wishart DS. Applications of metabolomics in drug discovery and development. Drugs in R & D. 2008;**9**(5):307-322. DOI: 10.2165/ 00126839-200809050-00002

[74] Cevallos-Cevallos JM, Reyes-De-Corcuera JI, Etxeberria E, Danyluk MD, Rodrick GE. Metabolomic analysis in food science: A review. Trends in Food Science & Technology. 2009;**20**(11):557-566. DOI: 10.1016/j. tifs.2009.07.002

[75] Mozzi F, Ortiz ME, Bleckwedel J, De Vuyst L, Pescuma M. Metabolomics as a tool for the comprehensive understanding of fermented and functional foods with lactic acid bacteria. Food Research International. 2013;**54**(1):1152-1161. DOI: 10.1016/j.foodres.2012.11.010

[76] Gupta A, Mann B, Kumar R, Sangwan RAMB. Antioxidant activity of Cheddar cheeses at different stages of ripening. International Journal of Dairy Technology. 2009;**62**(3):339-347. DOI: 10.1111/j.1471-0307.2009.00509.x

*Cheese Ripening: Impact on Cheese as a Functional Food DOI: http://dx.doi.org/10.5772/intechopen.114059*

[77] Pritchard SR, Phillips M, Kailasapathy K. Identification of bioactive peptides in commercial Cheddar cheese. Food Research International. 2010;**43**(5):1545-1548. DOI: 10.1016/j.foodres.2010.03.007

[78] Silva RA, Lima MSF, Viana JBM, Bezerra VS, Pimentel MCB, Porto ALF, et al. Can artisanal "Coalho" cheese from northeastern Brazil be used as a functional food? Food Chemistry. 2012;**135**(3):1533-1538. DOI: 10.1016/j. foodchem.2012.06.058

[79] Paul M, Brewster J, Hekken D, Tomasula P. Measuring the antioxidant activities of queso Fresco after post-packaging high pressure processing. Advances in Bioscience and Biotechnology. 2012;**03**:297-303. DOI: 10.4236/abb.2012.34042

[80] Bottesini C, Paolella S, Lambertini F, Galaverna G, Tedeschi T, Dossena A, et al. Antioxidant capacity of water soluble extracts from Parmigiano-Reggiano cheese. International Journal of Food Sciences and Nutrition. 2013;**64**(8):953-958. DOI: 10.3109/09637486.2013.821696

[81] Abadía-García L, Cardador A, Martín del Campo ST, Arvízu SM, Castaño-Tostado E, Regalado-González C, et al. Influence of probiotic strains added to cottage cheese on generation of potentially antioxidant peptides, anti-listerial activity, and survival of probiotic microorganisms in simulated gastrointestinal conditions. International Dairy Journal. 2013;**33**(2):191-197. DOI: 10.1016/j. idairyj.2013.04.005

[82] Smacchi E, Gobbetti M. Peptides from several Italian cheeses inhibitory to proteolytic enzymes of lactic acid bacteria, pseudomonas fluorescens ATCC 948 and to the angiotensin I-converting enzyme. Enzyme and Microbial

Technology. 1998;**22**(8):687-694. DOI: 10.1016/S0141-0229(97)00261-5

[83] Okamoto A, Hanagata H, Matsumoto E, Kawamura Y, Koizumi Y, Yanagida F. Angiotensin I converting enzyme inhibitory activities of various fermented foods. Bioscience, Biotechnology, and Biochemistry. 1995;**59**(6):1147-1149. DOI: 10.1271/ bbb.59.1147

[84] Saito T, Nakamura T, Kitazawa H, Kawai Y, Itoh T. Isolation and structural analysis of antihypertensive peptides that exist naturally in gouda cheese. Journal of Dairy Science. 2000;**83**(7):1434-1440. DOI: 10.3168/jds.S0022-0302(00)75013-2

[85] Gómez-Ruiz JÁ, Taborda G, Amigo L, Recio I, Ramos M. Identification of ACE-inhibitory peptides in different Spanish cheeses by tandem mass spectrometry. European Food Research and Technology. 2006;**223**:595- 601. DOI: 10.1007/s00217-005-0238-0

[86] Diana M, Rafecas M, Arco C, Quilez J. Free amino acid profile of Spanish artisanal cheeses: Importance of gammaaminobutyric acid (GABA) and ornithine content. Journal of Food Composition and Analysis. 2014;**35**:94-100. DOI: 10.1016/j. jfca.2014.06.007

[87] Siragusa S, De Angelis M, Di Cagno R, Rizzello C, Coda R, Gobbetti M. Synthesis of -aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses. Applied and Environmental Microbiology. 2007;**73**:7283-7290. DOI: 10.1128/AEM.01064-07

[88] Saidi V, Sheikh-Zeinoddin M, Kobarfard F, Soleimanian-Zad S. Bioactive characteristics of a semi-hard non-starter culture cheese made from raw or pasteurized sheep's milk. 3 Biotech. 2020;**10**(3):85. DOI: 10.1007/ s13205-020-2075-z

[89] Zeppa G, Conterno L, Gerbi V. Determination of organic acids, sugars, diacetyl, and acetoin in cheese by highperformance liquid chromatography. Journal of Agricultural and Food Chemistry. 2001;**49**(6):2722-2726. DOI: 10.1021/jf0009403

[90] Murtaza M, Rehman S, Anjum F, Huma N, Tarar O, Mueen-Ud-Din G. Organic acid contents of buffalo milk cheddar cheese as influenced by accelerated ripening and sodium salt. Journal of Food Biochemistry. 2012;**36**:99-106. DOI: 10.1111/j.1745-4514.2010.00517.x

[91] Jalilzadeh A, Tuncturk Y, Hesari J. Extension shelf life of cheese: A review. International Journal of Dairy Science. 2015;**10**:44-60. DOI: 10.3923/ ijds.2015.44.60

[92] Młynek K, Oler A, Zielińska K, Tkaczuk J, Zawadzka W. The effect of selected components of milk and ripening time on the development of the hardness and melting properties of cheese. Acta Scientiarum Polonorum. Technologia Alimentaria. 2018;**17**(2):133- 140. DOI: 10.17306/J.AFS.0549

[93] Delves-Broughton J. Natural antimicrobials as additives and ingredients for the preservation of foods and beverages. In: Natural Food Additives, Ingredients and Flavourings. Cambridge, UK: Elsevier; 2012. pp. 127-161

[94] Verheul A, Russell NJ, Van'T Hof R, Rombouts FM, Abee T. Modifications of membrane phospholipid composition in nisin-resistant *listeria monocytogenes* Scott A. Applied and Environmental Microbiology. 1997;**63**(9):3451-3457. DOI: 10.1128/aem.63.9.3451-3457.1997

[95] Benech R-O, Kheadr EE, Laridi R, Lacroix C, Fliss I. Inhibition of Listeria innocua in cheddar cheese by addition of nisin Z in liposomes or by in situ production in mixed culture. Applied and Environmental Microbiology. 2002;**68**(8):3683-3690. DOI: 10.1128/ AEM.68.8.3683-3690.2002

[96] Hassan H, St-Gelais D, Gomaa A, Fliss I. Impact of nisin and nisinproducing *Lactococcus lactis* ssp. lactis on clostridium tyrobutyricum and bacterial ecosystem of cheese matrices. Foods. Basel, Switzerland; 2021;**10**(4). DOI: 10.3390/foods10040898

[97] Moatsou G, Moschopoulou E, Beka A, Tsermoula P, Pratsis D. Effect of natamycin-containing coating on the evolution of biochemical and microbiological parameters during the ripening and storage of ovine hard-Gruyère-type cheese. International Dairy Journal. 2015;**50**:1-8. DOI: 10.1016/j. idairyj.2015.05.010

[98] Hashemi MM, Aminlari M, Forouzan MM, Moghimi E, Tavana M, Shekarforoush S, et al. Production and application of lysozyme-gum Arabic conjugate in mayonnaise as a natural preservative and emulsifier. Polish Journal of Food and Nutrition Sciences. 2018;**68**(1):33-43. DOI: 10.1515/ pjfns-2017-0011

[99] Nájera AI, Nieto S, Barron LJR, Albisu M. A review of the preservation of hard and semi-hard cheeses: Quality and safety. International Journal of Environmental Research and Public Health. Basel, Switzerland; 2021;**18**(18). DOI: 10.3390/ijerph18189789

[100] Brasca M, Morandi S, Silvetti T, Rosi V, Cattaneo S, Pellegrino L. Different analytical approaches in assessing antibacterial activity and the purity of commercial lysozyme preparations for dairy application. Molecules (Basel, Switzerland).

*Cheese Ripening: Impact on Cheese as a Functional Food DOI: http://dx.doi.org/10.5772/intechopen.114059*

2013;**18**(5):6008-6020. DOI: 10.3390/ molecules18056008

[101] El Soda M, Pandian S. Recent developments in accelerated cheese ripening. Journal of Dairy Science. 1991;**74**(7):2317-2335. DOI: 10.3168/jds. S0022-0302(91)78405-1

[102] Alhelli AM, Mohammed NK, Khalil ES, Hussin ASM. Optimizing the acceleration of Cheddar cheese ripening using response surface methodology by microbial protease without altering its quality features. AMB Express. 2021;**11**(1):45. DOI: 10.1186/ s13568-021-01205-9

#### **Chapter 3**

## Cheese's Bioactive Peptide Content and Fatty Acids Profile

*Ilyes Dammak and Carlos A. Conte-Junior*

#### **Abstract**

This chapter provides an in-depth review of the latest research developments in cheese's bioactive peptides and fatty acid profiles, emphasizing their potential health benefits, particularly in managing obesity and hyperlipidemia. It delves into the generation of bioactive peptides during cheese fermentation and maturation, their potential health-promoting effects, and the factors influencing their content. The chapter also offers a comprehensive analysis of the fatty acid profile in cheese, discussing the impact of various cheese-making processes on this profile and the subsequent implications for human health. Furthermore, it explores innovative strategies for enhancing the bioactive peptide content and optimizing the fatty acid profile in cheese. These strategies include using bioactive edible films, which have shown promise in improving the microbial quality of cheese and reducing lipid oxidation, thereby extending its shelf life. The chapter also investigates the encapsulation of bioactive compounds, a technique that has been used to enhance the stability and functionality of these compounds. Through this comprehensive review, the chapter offers valuable insights into the potential of cheese as a source of health-promoting bioactive peptides and fatty acids and the various strategies for optimizing their content and functionality.

**Keywords:** bioactive peptides, fatty acids, cheese, health benefits, cheese processing

#### **1. Introduction**

Cheese, an extensively consumed dairy product, is highly regarded for its abundant flavor, pleasing texture, and significant nutritional content. The above substance is a notable reservoir of proteins, fats, vitamins, and minerals. Cheese is not only rich in conventional nutrients, but it also contains bioactive peptides and a distinctive composition of fatty acids. These components have garnered significant attention in scientific research due to their potential positive effects on human health [1, 2].

Bioactive peptides are specific segments of proteins that have advantageous effects on different physiological functions or states, potentially influencing overall wellbeing. The peptides are encoded within the primary structure of the parent protein and can potentially be released through various mechanisms, such as digestion or food processing. Peptide formation is a prevalent process observed in cheese production, primarily facilitated by fermentation and maturation. Recent studies have

provided evidence suggesting that bioactive peptides can significantly impact health conditions such as obesity and hyperlipidemia [3, 4].

In contrast, the fatty acid composition in cheese is primarily determined by factors such as the origin of the milk, the feeding habits of dairy animals, and the methodologies utilized during the cheese manufacturing processes. The association of various health benefits with fatty acids, particularly unsaturated ones, has been documented [5, 6].

In addition to these naturally occurring compounds, innovative strategies are being developed to preserve cheese's bioactive peptide content and fatty acid profile. One such strategy is the use of bioactive edible films, which have shown promise in improving the microbial quality of cheese and reducing lipid oxidation, thereby extending its shelf life [7, 8]. Another promising approach is the encapsulation of bioactive compounds, a technique that has been used to improve the stability and functionality of these compounds [9].

The cheese's bioactive peptides are influenced by the cheese-making process and the type of milk it produces. An investigation on goat milk cheese revealed a higher concentration of bioactive peptides than cow milk cheese [10]. Moreover, certain probiotic strains in cheese-making have been shown to enhance the bioactive peptide content. A study on cheddar cheese showed that using *Lactobacillus casei* 300 resulted in the formation of peptides with antihypertensive activity [11]. Moreover, the utilization of specific enzymes, such as proteases, in the cheese production process has demonstrated the ability to augment the presence of bioactive peptides. The investigation conducted on Gouda cheese demonstrated that utilizing proteases derived from *Lactobacillus helveticus* led to the production of peptides exhibiting antihypertensive properties [12]. In addition, a study on blue cheese showed that the ripening process led to the formation of peptides with antioxidant activity [13]. Furthermore, a study on Camembert cheese showed that the ripening process led to the formation of peptides with antithrombotic activity [14]. Finally, a study on Roquefort cheese showed that the ripening process led to the formation of peptides with anti-inflammatory activity [15].

For cheese processing, reducing salt content in cheese through methods such as decreasing the brine soaking time has not impacted the bioactive peptide formation or fatty acid bioaccessibility in cheese [15]. The ripening process of cheese also plays a significant role in forming bioactive peptides. For instance, a study on Mexican goat cheese showed that the ripening process led to the formation of peptides with antioxidant activity [13]. Furthermore, an investigation conducted on Dutch-style cheese revealed that the presence of fatty acids and conjugated fatty acids not only plays a role in the overall nutritional profile of cheese but also, when combined with chemometric techniques, can serve as chemical biomarkers for evaluating the source and maturation status of cheeses, as well as verifying their authenticity [8]. Additionally, a research investigation on Parmigiano–Reggiano cheese revealed that the defatting procedure could retain the entire cheese's nutritional characteristics, encompassing bioactive peptides. This was accomplished through high-performance liquid chromatography (HPLC) [16].

The incorporation of plant-based diets, such as flaxseed, in dairy animals has been demonstrated to enhance the nutritional profile of cheese [10, 11]. Flaxseed is recognized as a substantial reservoir of α-linolenic acid, classified as an omega-3 fatty acid, and widely acknowledged for its advantageous impact on cardiovascular well-being. Research has indicated that the inclusion of flaxseed in the diet of dairy animals results in an elevated presence of omega-3 fatty acids in their milk. Consequently,

*Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

this dietary modification contributes to the resulting cheese's enhanced fatty acid composition [10, 11].

Similarly, certain forages in dairy animals' diets can enhance cheese's bioactive peptide content. A notable example is sulla, a leguminous forage, which has been shown to increase the bioactive peptide content in cheese [17]. Bioactive peptides refer to fragments of proteins that can positively impact bodily functions or conditions, potentially influencing overall health. Moreover, the germination of certain seeds, such as black soybeans, has been found to enhance their anti-Alzheimer activity [14]. This suggests a potential application in cheese production, where these germinated seeds could be used as an ingredient or supplement. The resultant cheese could have enhanced neuroprotective properties, offering a novel approach to preventing or managing neurodegenerative diseases like Alzheimer's. Later, dairy animals' diet and specific ingredients or supplements can significantly influence cheese's nutritional and bioactive properties. This opens up exciting possibilities for the production of functionally enhanced cheese products, contributing to the field of functional foods.

This chapter aims to comprehensively review recent research developments on cheese's bioactive peptides and fatty acid profiles. It will delve into their healthpromoting potential, the factors influencing their content, and the innovative strategies for enhancing their presence and functionality in cheese. The chapter is designed to be accessible to readers from various fields, not just those who are dairy science or nutrition experts.

#### **2. Bioactive peptides in cheese**

In the realm of functional foods, cheese holds a unique position due to its rich content of bioactive peptides and specific fatty acid profiles. This section delves into the intricate processes that lead to the generation of these bioactive peptides during cheese production and the subsequent health benefits they confer.

#### **2.1 Generation of bioactive peptides in cheese**

Producing bioactive peptides in cheese is a complex procedure that necessitates a comprehensive comprehension of the biochemical and microbiological mechanisms at play for its thorough analysis. Hence, acquiring a thorough comprehension of these factors can facilitate the optimization of the proteolysis process and augment the production of bioactive peptides. These peptides are primarily produced due to gastrointestinal digestion, a multifaceted physiological process characterized by the enzymatic degradation of proteins into smaller peptides and amino acids. Simultaneously, the process of milk processing, which involves pasteurization and homogenization, has the potential to generate these bioactive peptides. Moreover, enzymatic hydrolysis, which involves the enzymatic degradation of proteins into smaller peptides, constitutes another noteworthy factor. Indeed, many digestive enzymes, including pepsin, trypsin, and chymotrypsin, are responsible for the cleavage of proteins at specific locations. Microbial fermentation, a biological process in which microorganisms such as bacteria and yeast metabolize organic substances, frequently produces peptides. The various processes, both independently and in combination, play a significant role in producing bioactive peptides. These peptides have garnered considerable attention due to their potential to promote health (**Figure 1**).

#### **Figure 1.** *Production mechanisms of bioactive peptides [18].*

The enzymatic degradation of proteins by starter cultures plays a crucial role in this process [19]. Nevertheless, it is crucial to consider the involvement of nonstarter cultures, as they can play a significant role in the proteolytic activity throughout the ripening phase and influence the cheese's ultimate peptide composition. Within the given framework, nonstarter cultures, called secondary or adjunct cultures, encompass bacteria deliberately introduced into the milk used for cheese production alongside the primary starter cultures. Nevertheless, their contribution to the acidification of the cheese curd, the primary function of starter cultures, is not substantial. In contrast, cultures that do not initiate fermentation play a significant role in shaping cheese's flavor, texture, and various attributes as it undergoes the ripening phase. Nonstarter cultures encompass various microorganisms, including lactobacilli, propionibacteria, and specific fungal species. These organisms can synthesize enzymes that catalyze the hydrolysis of proteins, fats, and carbohydrates in cheese, forming flavor compounds and other substances, such as bioactive peptides [20].

The research conducted by Kurbanova et al. offers significant insights into the contribution of distinct starter cultures in producing bioactive peptides [21]. However, it would be interesting to investigate further the potential influence of the interaction between various starter and nonstarter cultures on the peptide profile. For example, what impact would the inclusion of additional strains of lactobacilli or other bacterial species have on the production of bioactive peptides? Furthermore, it would be advantageous to investigate the impact of fermentation conditions, including temperature and duration, on the efficacy of these cultures and the consequent production of peptides.

The significance of proteolysis in milk conversion into cheese, as emphasized by Lepilkina and Grigorieva [22], is undeniably pivotal. Nevertheless, it is important to acknowledge that proteolysis is a multifaceted phenomenon that can be affected by various factors, such as the specific enzymes utilized, the properties of the milk proteins, and the parameters of the cheese production procedure. Hence, acquiring a thorough comprehension of these factors can facilitate the optimization of the proteolysis process and augment the production of bioactive peptides.

#### *Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

The research conducted by Helal et al. [23] offers a noteworthy viewpoint on the temporal aspects of proteolysis and the production of bioactive peptides in whey fermentation. However, further investigation into optimizing the fermentation regime is warranted to maximize the production of specific bioactive peptides. For example, could an extended duration of fermentation lead to an increased concentration of specific peptides? Alternatively, could it result in the deterioration of these peptides and the formation of additional compounds?

The study conducted by Araújo-Rodrigues et al. highlights the potential of utilizing autochthonous starter cultures to improve cheese's sensory attributes and safety characteristics [24]. Nevertheless, it is crucial to consider the potential obstacles linked to this methodology, including the fluctuation in microbial populations within unpasteurized milk and the possible impact of environmental factors on the functionality of these microorganisms.

Furthermore, it is crucial to emphasize the research conducted by Sturova et al. [25], which involved the development of a distinctive approach to cheese maturation. This method involved the utilization of a noble mold derived from a combination of whole milk and secondary protein-carbohydrate raw materials. Their research findings indicated that the proteolysis and lipolysis processes exhibited higher-intensity levels in the experimental cheeses manufactured using a noble mold. Consequently, the final product displayed enhanced organoleptic qualities. Utilizing this methodology can potentially augment the production of bioactive peptides within the context of cheese manufacturing.

In addition, Samelis et al. conducted a study to assess the efficacy of a blended thermophilic and mesophilic starter culture comprising *Streptococcus thermophilus* ST1 and the Greek autochthonous nisin-A-producing *Lactococcus lactis* in the production of the traditional Galotyri Protected Designation of Origin (PDO) cheese [26]. According to their study, it has been indicated that the distinctive characteristics of cheese, including its fermentation techniques and the type of milk employed, have the potential to affect the behavior of bacteria and the subsequent production of bioactive peptides.

In addition, a research study conducted by Moiseenko et al. examined the capacity of *Lacticaseibacillus paracasei* strains, which were obtained from the traditional South African fermented beverage mahewu and kefir grains, to generate bioactive peptides possessing antioxidant and angiotensin I-converting enzyme inhibitory (ACE-I) characteristics during the process of milk fermentation [27]. The current research study provides valuable insights into the exploration of unconventional isolation sources for synthesizing bioactive peptides in the domain of cheese production. To summarize, the generation of bioactive peptides in cheese is a complex occurrence that is influenced by various factors, including the use of diverse starter and nonstarter cultures, the intricacies of the fermentation process, and the unique characteristics specific to the type of cheese being produced. In order to optimize the generation of bioactive peptides and enhance the health-promoting properties of cheese, future investigations should prioritize a thorough analysis of these factors.

In summary, the generation of bioactive peptides in cheese is a complex process influenced by various factors, including the use of diverse starter and nonstarter cultures, the intricacies of the fermentation process, and the unique characteristics specific to the type of cheese being produced. Understanding these factors can help optimize the proteolysis process and enhance the production of bioactive peptides.

Transitioning to the health benefits of these peptides, it is important to note that the bioactive peptides produced in cheese have been associated with various healthpromoting effects.

#### **2.2 Health-promoting effects of bioactive peptides in cheese**

Bioactive peptides in cheese have been associated with various health benefits. These peptides are thought to exert their beneficial effects through various mechanisms, including inhibiting enzymes involved in disease processes, modulation of the immune system, and scavenging free radicals (**Table 1**). These studies present supplementary evidence to substantiate that cheese can produce bioactive peptides that confer various health benefits, including antithrombotic and antidiabetic properties.

In a recent study conducted by Helal et al. [23], the researchers examined six distinct types of cheese to assess the concentrations of bioactive peptides and their corresponding physiological effects. The cheeses examined in this study consisted of three distinct types of Egyptian cheese, namely Karish, Domiati, and Ras, and three internationally popular cheese varieties, including Feta-type, Gouda, and Edam. The study's results revealed that a significant portion of the bioactive peptides identified exhibited inhibitory effects on crucial enzymes associated with the advancement of cardiovascular diseases, specifically angiotensin-converting enzyme (ACE), as well as diabetes, particularly dipeptidyl peptidase-IV (DPP-IV). It is worth mentioning that Gouda cheese exhibited the most significant ACE inhibitory and DPP-IV-inhibitory activities, in addition to displaying the highest level of antioxidant activity. Further investigation has clarified the involvement of bioactive peptides in the composition of cheese and other edible substances. The study conducted by Helal et al. [28] contributes additional knowledge to the ongoing scholarly conversation regarding synthesizing bioactive peptides in cheese. The primary objective of this study was to investigate the effects of spontaneous fermentation and the inclusion of natural whey starters on the peptidomics profile and biological activities of cheese whey. The study revealed spontaneous fermentation incorporating natural whey starters considerably deteriorated whey proteins. The breakdown process facilitated the subsequent release of peptides with biological activity and enhanced the digestibility of the protein content. The researchers conducted an investigation involving identifying more than four hundred peptides. The peptides being examined were primarily derived from β-casein, κ-casein, and α-lactalbumin. Out of the identified peptides, a collective count of 49 exhibited bioactive characteristics, with 21 of these peptides demonstrating inhibitory effects on the angiotensin-converting enzyme (ACE).


#### **Table 1.**

*Biologically active peptides isolated from several cheeses.*

#### *Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

Moreover, the study additionally revealed that the lactotripeptide isoleucineproline-proline (IPP) exhibited higher levels compared to valine-proline-proline (VPP), with the most significant concentration observed during the period of spontaneous fermentation after 24 hours. The results of this study indicate that the fermentation process, regardless of whether it occurs spontaneously or is initiated using natural whey starters, plays a significant role in producing bioactive peptides in cheese. Rehman et al. [29] conducted an independent investigation to evaluate the therapeutic efficacy of water-soluble peptides (WSPs) derived from probiotic cheddar cheese produced from buffalo milk. The study focused specifically on the antithrombotic properties of these peptides [29]. The study's results demonstrate a significant increase in antithrombotic activity as the ripening period progressed for both the control and probiotic cheddar cheese samples.

Incorporating a probiotic adjunct in cheddar cheese production led to a significant improvement in its antithrombotic activity. Pontonio et al. conducted an independent study to increase the worth of whey obtained from ricotta cheese. Their approach involved the development of a biotechnological method for generating bioactive peptides that demonstrate inhibitory properties against angiotensin-Iconverting enzyme (ACE) [30]. The study utilized a methodology that incorporated the combination of membrane filtration and fermentation techniques. The fermented R-UF demonstrated a notable anti-angiotensin-converting enzyme (ACE) activity. The purified active fractions of the fermented R-UF displayed sequences that partially or fully coincided with κ-casein antihypertensive fragments that have been previously reported. The ricotta cheese, which was enriched with a fortification level of 5%, demonstrated a concentration of approximately 30 mg of bioactive peptides. The investigation conducted by Vázquez-García et al. involved the assessment of peptide fractions found in Mexican goat cheeses during different ripening periods [13]. An examination was conducted to explore the correlation between the peptide fractions and the antioxidative properties demonstrated by the cheeses [13]. The study findings indicate that the observed DPPH radical scavenging activity in ripened Mexican goat cheese can be attributed to the peptides that naturally occur in the milk or are produced as a result of the action of starter cultures during the cheese ripening process.

In addition, Martini et al. undertook a research investigation aimed at assessing the impact of different stages of ripening in Parmigiano–Reggiano (PR) cheese peptide fractions on the enzymatic activity of α-glucosidase, α-amylase, and dipeptidyl peptidase-IV (DPP-IV), as well as the formation of fluorescent advanced glycation end-products (fAGEs) [31]. The PR peptide fractions exhibited inhibitory properties against the specific enzymes and the formation of fAGEs. These studies present supplementary evidence to substantiate that cheese can produce bioactive peptides that confer various health benefits, including antithrombotic and antidiabetic properties. Bioactive peptides demonstrate diverse pharmacological characteristics, encompassing opioid activity, anti-tumor effects, anti-lipidemic properties, and immunomodulatory activities. As depicted by Shafique et al. [32], cheddar cheese-derived bioactive peptides have the potential as functional foods, affecting chronic diseases like obesity, cardiovascular, and diabetes. These peptides have multifunctional therapeutic potentials, including antimicrobial, immunomodulatory, antioxidant, enzyme inhibitory, antithrombotic, and phytopathological effects. They regulate immune, gastrointestinal, hormonal, and neurological responses, which are crucial in disease prevention and treatment. **Figure 2** illustrates the depiction of the action mechanism of bioactive peptides.

**Figure 2.** *Action mechanism of bioactive peptides [32].*

In summary, bioactive peptides in cheese have been linked to a range of health benefits, including antithrombotic and antidiabetic properties. These peptides have multifunctional therapeutic potentials, including antimicrobial, immunomodulatory, antioxidant, enzyme inhibitory, antithrombotic, and phytopathological effects. They regulate immune, gastrointestinal, hormonal, and neurological responses, which are crucial in disease prevention and treatment.

In the next section, we will delve into the fatty acid profile of cheese and how it contributes to the overall nutritional value and health benefits of this popular dairy product.

#### **3. Fatty acid profile in cheese and its health implications**

The fatty acid composition of cheese is primarily influenced by the origin of the milk, as well as the feeding practices of dairy animals and the methods employed during the cheese manufacturing process. The consumption of unsaturated fatty acids has been linked to numerous health advantages. Gas chromatography (GC) is a widely recognized analytical method that provides high precision and accuracy in the quantification of fatty acid composition in cheese. The findings are commonly reported in milligrams of fatty acids per gram of cheese (mg FA/g cheese), as illustrated in **Table 2**. The obtained data comprehensively analyze the fatty acid composition of the cheese, encompassing saturated, monounsaturated, and polyunsaturated fatty acids. The provision of this information is of utmost importance in comprehending the nutritional characteristics of cheese, as distinct fatty acids exert varying impacts on human well-being. For instance, it is widely acknowledged in the academic literature


#### **Table 2.**

*Cheese's fatty acid profiles (mg FA/g cheese) as determined by gas chromatography [33].*

that monounsaturated and polyunsaturated fatty acids are generally regarded as advantageous for maintaining cardiovascular health.

The fatty acid composition of cheese exhibited notable variations throughout the production season, potentially influenced by many factors, including the grazing patterns of cows, moisture content in their feed, ambient temperature, lactation stage, and the overall health condition of the animals [34].

Recent studies have investigated dietary factors' influence on dairy products' fatty acid composition. For example, a research investigation on Kivircik ewes revealed that the incorporation of 3% palm oil into their dietary regimen resulted in a notable elevation in the concentration of palmitic acid in their milk, escalating from 28% to 36% [35]. The observed rise in palmitic acid concentration in the milk corresponds to a 29% increase, assuming that the palm oil utilized contained 80% palmitic acid. The research study additionally demonstrated that the incorporation of palm oil into the dietary regimen resulted in elevated levels of saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA), primarily attributable to heightened concentrations of palmitic acid (C16:0) and oleic acid (C18:1), respectively.

The ramifications for human health resulting from alterations in fatty acid composition are multifaceted and intricate. Based on a specific perspective, there exists evidence indicating a potential association between certain saturated fatty acids, namely lauric (C12:0) and myristic (C14:0) acids, and a decreased risk of developing coronary heart disease [36]. On the other hand, incorporating palm oil into one's dietary intake leads to an elevation in the collective levels of saturated fatty acids, which have been established to have adverse effects on human well-being [37].

Their fatty acid profile determines the nutritional quality of dairy products, and various indices are employed to evaluate the diet's nutritional value and its impact on consumer health. The research presented empirical findings that the inclusion of 3% rumen-protected palm oil in the dietary regimen enhanced ewe milk's health characteristics, particularly in relation to the optimal levels of desirable fatty acids (DFA) for the n-6/n-3 ratio. Despite the lack of observed improvement in the atherogenicity index (AI), thrombogenicity index (TI), health-promoting index (HPI), and h/H (hypocholesterolemic/hypercholesterolemic acids) indices, the values obtained in this study align with those documented in prior scholarly works. This discovery suggests that the milk examined in the current investigation does not harm consumers' health [35].

The research conducted by Zajác et al. examined the fatty acid composition of traditional Slovak cow's lump cheese, as depicted in **Figure 3** [38]. The cheese, which a nearby local farmer manufactures, exhibited diverse levels of fatty acids. The primary fatty acids identified in cow's lump cheese were palmitic acid (C16:0) with a concentration range of 29–37 g/100 g, oleic acid (C18:1) (n-9) with a concentration range of 19–26 g/100 g, myristic acid (C14:0) with a concentration range of 10–13 g/100 g, and stearic acid (C18:0) with a concentration range of 8–10 g/100 g. The investigation recorded noteworthy alterations in the concentrations of particular fatty acids throughout the production duration. The research determined that the fat content of cow's lump cheese was 23%, with a standard deviation of ±1.5%. The lowest observed fat content was 21%, whereas the highest recorded fat content was 27%. The cheese

**Figure 3.** *Cow's lump cheese combines saturated, monounsaturated, and polyunsaturated fatty acids [38].*

#### *Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

sample exhibited a composition of 70 g/100 g for saturated fatty acids, 26 g/100 g for monounsaturated fatty acids, and 3 g/100 g for polyunsaturated fatty acids.

The sensory attributes of cheese, including color, texture, flavor, and nutritional value, are notably impacted by the composition of fatty acids in the cheese. Various types of fatty acids have the potential to impact human health, with certain ones offering benefits while others may pose potential risks. The fatty acid composition can also function as a discriminative characteristic for different types of cheese, depending on the milk utilized during their manufacturing process. In their investigation, González-Martín et al. utilized Near-Infrared Spectroscopy (NIRS) technology to determine the presence of 19 fatty acids in cheese, ranging from C8:0 to C20:0. The evaluation involved the analysis of both the aggregate amount of saturated fatty acids (∑SFA) and the aggregate amount of unsaturated fatty acids (∑UFA). The research study substantiated the feasibility of Near-Infrared Spectroscopy (NIRS) as a prompt, dependable, and effective technique for obtaining data on the lipid composition of cheese samples [33].

Furthermore, it is important to exercise caution when interpreting the use efficiencies of diet ingredients in production systems that employ varying ingredient inclusion rates. The organic production systems demonstrated higher efficiency in nongrazing and concentrated utilization, underscoring the significance of incorporating pasture in cows' diets. This approach reduces the reliance on costly ingredients that compete with human food sources and presents opportunities to enhance profitability and sustainability within the system by promoting greater pasture intake. The existing data do not provide sufficient evidence to support the notion that incorporating higher levels of concentrate or nonpasture feeds in the diets of organic cows is justified. This is because the data does not demonstrate that organic herds possess a greater capacity to utilize concentrate or nonpasture ingredients more effectively than conventional herds.

The current study's findings suggest that organic herds demonstrated a reduced occurrence of mastitis cases, expressed as a proportion of the overall herd, compared to conventional herds. This observation is consistent with the findings documented by Ellis et al. in their study [39]. In contrast, Stergiadis et al. [40] observed no significant variation in mastitis occurrences across herds with varying levels of production intensity, ranging from organic to highly intensive. Nevertheless, in terms of numerical data, it was observed that the higher-intensity systems exhibited a greater incidence of mastitis cases, even in the presence of preventive antibiotic measures. Nevertheless, the Redundancy Analysis (RDA), as determined in the present study, aligns with the findings of Stergiadis et al., as it reveals the presence of adverse correlations between grazing practices and incidences of mastitis [40]. Hence, the elevated levels of pasture consumption within organic systems may have limited significance for mastitis. In their study, Ellis et al. [39] and Ward et al. observed a correlation between enhanced cow cleanliness on farms and decreased mastitis cases and somatic cell count (SCC). However, the authors reported no significant disparity in cow cleanliness during outdoor grazing between organic and conventional systems [41]. The study provides no documentation regarding the cleaning and milking strategies employed.

Consequently, it is not feasible to make any assertions regarding the potential influence of these strategies on mastitis cases. Nevertheless, previous research has established a correlation between genetic selection for a high milk yield, particularly observed in the Holstein breed, which was more prevalently utilized in the conventional herds examined in this current study, and somatic cell count (SCC) levels,

consequently leading to mastitis [42]. The present study observed that the organic herds exhibited reduced milk yields and somatic cell counts (SCC), alongside a greater proportion of lower-yielding breeds such as Ayrshire and Shorthorn. This particular breed composition could potentially account for the decreased incidence of mastitis.

Previous research has demonstrated that low-input and organic milk display reduced levels of saturated fatty acids (SFA) compared to high-intensity and conventional milk [43, 44]. In contrast to prior research, the current study reveals divergent outcomes. Prior studies have demonstrated a reduced concentration of saturated fatty acids (SFAs) in organic milk compared to milk derived from intensively managed herds [45]. The observed disparity has been ascribed to the elevated intake of fresh herbage within organic farming systems. However, no notable differences were observed between the organic and conventional systems regarding pasture intake. The variation in intake was minimal, measuring 123 g/kg dry matter (DM) [46]. The present study reveals a comparable pattern, wherein the disparity in the proportion of total forage and pasture between the conventional and organic herds was found to be less than 131 g/kg and 166 g/kg DM per day, respectively. The results derived from the RDA demonstrate a significant association between the consumption of whole crops and grass silage and the saturated fatty acid (SFA) content of milk compared to grazing. As mentioned earlier, the observation is supported by the research conducted by Ellis et al., wherein their investigation revealed that incorporating whole crops and grass silage into the diet increased saturated fatty acids (SFA) levels in milk [47].

Furthermore, Ormston et al. [48] discovered a positive correlation between saturated fatty acid (SFA) consumption and grass and maize silage intake. The observed disparity in organic diets, with average values of 778 g/kg DM and 920 g/kg DM for whole crop and grass silage, respectively, compared to conventional diets, may have played a significant role in the elevated levels of saturated fatty acids (SFA) found in organic milk during the present investigation. Furthermore, substantial evidence supports that breed is crucial in determining the fatty acid (FA) profile. This is evident from studies that have observed lower concentrations of saturated fatty acids (SFA) in Holstein-Friesian cows' milk than in other breeds [48, 49]. The present study's findings also demonstrated a positive association between certain individual fatty acids (C6:0, C8:0, C10:0, C12:0, and C14:0) and non-Holstein breeds, as indicated by the results obtained from RDA. However, the disparities in the breed systems were not found to be statistically significant. Conventional herds exhibited a slight numerical advantage of approximately 14 Holstein cows per 100 cows compared to crossbreeds and organic herds. The observed variation in the composition of the herd may have played a role in the observed rise in the levels of specific saturated fatty acids (SFAs) in the milk, including C6:0, C8:0, and C14:0. It is worth mentioning that the somatic cell count (SCC) in organic milk exhibited a higher value exclusively during September to December, a period characterized by minimal or zero pasture intake in both agricultural systems. This finding is consistent with the research conducted by Butler et al. [43], which observed a higher level of saturated fatty acids (SFA) in milk from organic farms compared to nonorganic low-input farms during the indoor period in August and October.

In summary, the fatty acid composition of cheese has complex and multifaceted implications for human health. While certain saturated fatty acids have been linked to an increased risk of cardiovascular disease, other fatty acids, such as monounsaturated and polyunsaturated fatty acids are generally regarded as beneficial for cardiovascular health. The fatty acid profile of cheese can also be influenced by

#### *Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

various factors, including the grazing patterns of cows, the moisture content in their feed, ambient temperature, lactation stage, and the overall health condition of the animals. Therefore, understanding these factors and their impact on the fatty acid profile of cheese is crucial for optimizing the health benefits of cheese consumption.

In the following section, we will delve into the innovative strategies aimed at enhancing the health benefits of cheese consumption. Specifically, we will focus on methods designed to increase the production of bioactive peptides and optimize the fatty acid profile in cheese, as these components play a crucial role in promoting health and preventing disease.

#### **4. Innovative strategies for enhancing bioactive peptides and optimizing fatty acid profile in cheese**

In conjunction with naturally existing compounds, novel methodologies are being devised to augment the bioactive peptide composition and optimize the fatty acid configuration in cheese. One strategy that has been explored is the utilization of bioactive edible films. These films have demonstrated the potential to enhance the microbial characteristics of cheese and mitigate lipid oxidation, consequently prolonging its shelf life [50, 51]. In cheese preservation, Aloe-based bioactive edible films have been observed to enhance the cheese's lipid stability and microbial quality (**Figure 4**). *Aloe vera*, a botanical species widely recognized for its therapeutic attributes, encompasses diverse bioactive constituents, such as polysaccharides, vitamins, enzymes, and antioxidants. Including these bioactive compounds in an edible film can provide advantages to the packaged food item [7]. The Aloe film has been found to contain bioactive compounds that can inhibit lipid oxidation, which significantly contributes to the degradation of cheese quality [7].

Consequently, lipids' stability is improved, maintaining the cheese's sensory attributes and nutritional composition throughout its shelf life. In addition, the antimicrobial properties of *Aloe vera* have been found to contribute to the preservation of the microbial quality of cheese [7]. The bioactive compounds present in the Aloe film exhibit inhibitory effects on spoilage microorganisms and pathogens, thereby effectively prolonging the cheese's shelf life and ensuring its safety for consumption.

Encapsulation of bioactive compounds is a technique employed to improve the stability and functionality of these compounds [52], presenting itself as a promising method. Pop et al. [53] thoroughly examined the encapsulation process for *Moringa oleifera* bioactive compounds in their study. The authors emphasized the significant potential of encapsulation techniques in facilitating the incorporation of bioactive molecules into various food products.

#### **Figure 4.**

*A bioactive edible film derived from Aloe improved the lipid stability and microbial quality of cheese [7].*

Furthermore, the utilization of whey proteins in cheese manufacturing is gaining traction owing to their advantageous effects on human health. Whey proteins are a significant provider of essential amino acids, including leucine, isoleucine, and valine, classified as branched-chain amino acids [50]. Rapidly digestible proteins offer older individuals enhanced nutritional advantages compared to casein and other protein sources. As a result, their extensive integration into clinical nutritional products has been observed [50].

Nevertheless, the general reception of whey protein-fortified products among consumers tends to be low owing to unfavorable flavor and aroma characteristics and negative mouth-feel attributes. These include the accumulation of mouthdrying, mouth-coating, chalky, metallic, and filming sensations associated with repeated consumption, as indicated by previous research [50]. Hence, further investigation is necessary to improve the sensory characteristics of these products, thereby augmenting consumer acceptance and appropriateness. Concerning the composition of fatty acids, the utilization of distinct starter cultures and adjuncts has the potential to yield cheese products that exhibit an elevated content of conjugated linoleic acid (CLA) and omega-3 fatty acids, which are known to possess health-promoting properties [45].

Furthermore, it should be noted that specific processing techniques, such as highpressure processing (HPP), have the potential to modify the fatty acid composition of cheese. The study conducted by Inácio et al. [54] proved that subjecting cheddar cheese to high-pressure processing (HPP) resulted in an elevation of unsaturated fatty acid levels, particularly oleic acid.

In summary, pursuing novel approaches to augment the bioactive peptide composition and optimize the fatty acid makeup in cheese represents a vibrant and auspicious realm of scholarly investigation. The abovementioned strategies are designed to enhance cheese's nutritional composition, prolong its shelf life, and enhance its sensory characteristics.

#### **5. Conclusions**

Exploring cheese as a source of bioactive peptides and fatty acids has opened new avenues in functional foods. This chapter has provided a comprehensive overview of the current knowledge and recent advancements in this area. Bioactive peptides generated during cheese fermentation and maturation have been associated with various health benefits, including managing obesity and hyperlipidemia. They exert their beneficial effects through various mechanisms, such as inhibiting enzymes involved in disease processes, modulating the immune system, and scavenging free radicals. The content of these peptides in cheese is influenced by several factors, including the type of milk used, the cheese-making process, and the specific strains of bacteria involved in fermentation.

The fatty acid profile of cheese, particularly the unsaturated fatty acids, also plays a crucial role in human health. The type of milk used, the diet of the dairy animals, and the cheese-making processes largely determine this profile. Recent research has shown that specific starter cultures and adjuncts can produce cheese with a higher content of health-promoting fatty acids, such as conjugated linoleic acid (CLA) and omega-3 fatty acids.

Innovative strategies are being developed to enhance the bioactive peptide content and optimize the fatty acid profile in cheese. These include the use of bioactive edible

*Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

films and the encapsulation of bioactive compounds, which have shown promise in improving the microbial quality of cheese, reducing lipid oxidation, extending its shelf life, and improving the stability and functionality of these compounds.

In conclusion, cheese, a staple food item consumed worldwide, holds significant potential as a source of health-promoting bioactive peptides and fatty acids. The ongoing research and development in this field are expected to produce cheese varieties with enhanced health benefits, thereby contributing to the growing functional foods market. However, further research is needed to fully understand these bioactive compounds' mechanisms of action and optimize the cheese-making processes for their production. The potential of these findings extends beyond the cheese industry, offering insights that could be applied to the broader field of food science and nutrition.

#### **Acknowledgements**

This work was financially supported by the Brazilian National Council for Scientific and Technological Development (CNPq) (102263/2022-1).

#### **Conflict of interest**

The authors declare no conflict of interest.

#### **Author details**

Ilyes Dammak1 \* and Carlos A. Conte-Junior2

1 University of Kairouan, Tunisia

2 Federal University of Rio de Janeiro, Brazil

\*Address all correspondence to: dammakilyes@hotmail.fr

© 2023 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### **References**

[1] Lin P, Di H, Li Z, Wang Y,

Zhou W, Huang S, et al. Light irradiation maintains the sensory quality, healthpromoting phytochemicals, and antioxidant capacity of post-harvest baby mustard. Journal of Food Science. 2022;**87**(1):112-123

[2] Punia H, Tokas J, Malik A, Sangwan S, Baloda S, Singh N, et al. Identification and detection of bioactive peptides in milk and dairy products: Remarks about agro-foods. Molecules. 2020;**25**(15):3328

[3] Song Y, Cai Q, Wang S, Li L, Wang Y, Zou S, et al. The ameliorative effect and mechanisms of Ruditapes philippinarum bioactive peptides on obesity and hyperlipidemia induced by a high-fat diet in mice. Nutrients. 2022;**14**(23):5066

[4] Wang H, Dong P, Liu X, Zhang Z, Li H, Li Y, et al. Active peptide AR-9 from Eupolyphaga sinensis reduces blood lipid and hepatic lipid accumulation by restoring gut flora and its metabolites in a high fat diet–induced hyperlipidemia rat. Frontiers in Pharmacology. 2022;**13**:918505

[5] Pajor F, Várkonyi D,

Dalmadi I, Pásztorné-Huszár K, Egerszegi I, Penksza K, et al. Changes in chemical composition and fatty acid profile of milk and cheese and sensory profile of milk via supplementation of goats' diet with marine algae. Animals. 2023;**13**(13):2152

[6] Wasilewski R, Kokoszyński D, Włodarczyk K. Fatty acid profile, health lipid indices, and sensory properties of meat from Pekin ducks of different origins. Animals. 2023;**13**(13):2066

[7] Kouser F, Kumar S, Bhat HF, Hassoun A, Bekhit AE-DA, Bhat ZF. Aloe barbadensis based bioactive edible film improved lipid stability and microbial quality of the cheese. Food. 2023;**12**(2):229

[8] Robalo J, Lopes M, Cardoso O, Sanches Silva A, Ramos F. Efficacy of whey protein film incorporated with Portuguese green tea (*Camellia sinensis* L.) extract for the preservation of latin-style fresh cheese. Food. 2022;**11**(8):1158

[9] Hu Y-K, Bai X-L, Yuan H, Zhang Y, Ayeni EA, Liao X. Polyphenolic glycosides from the fruits extract of *Lycium ruthenicum* Murr and their monoamine oxidase B inhibitory and neuroprotective activities. Journal of Agricultural and Food Chemistry. 2022;**70**(26):7968-7980

[10] Dokou S, Athanasoulas A, Vasilopoulos S, Basdagianni Z, Dovolou E, Nanas I, et al. Composition, organoleptic characteristics, fatty acid profile and oxidative status of cow's milk and white cheese after dietary partial replacement of soybean meal with flaxseed and lupin. Animals. 2023;**13**(7):1159

[11] Giosuè C, D'Agostino F, Maniaci G, Avellone G, Sciortino M, De Caro V, et al. Persistent organic pollutants and fatty acid profile in a typical cheese from extensive farms: First assessment of human exposure by dietary intake. Animals. 2022;**12**(24):3476

[12] Chen Y, Liu W, Xue J, Yang J, Chen X, Shao Y, et al. Angiotensin-converting enzyme inhibitory activity of *Lactobacillus helveticus* strains from traditional fermented dairy foods and antihypertensive effect of fermented milk of strain H9. Journal of Dairy Science. 2014;**97**(11):6680-6692

*Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

[13] Vázquez-García R, Cardador-Martinez A, Orihuela-López MA, Ramos-Hernández LS, Martíndel-Campo ST. Preliminary study of extended ripening effects on peptides evolution and DPPH radical scavenging activity in Mexican goat cheese. Catalysts. 2021;**11**(8):967

[14] Shabbir U, Tyagi A, Ham HJ, Oh D-H. Comprehensive profiling of bioactive compounds in germinated black soybeans via UHPLC-ESI-QTOF-MS/MS and their anti-Alzheimer's activity. PLoS One. 2022;**17**(1):e0263274

[15] Di Nunzio M, Loffi C, Chiarello E, Dellafiora L, Picone G, Antonelli G, et al. Impact of a shorter brine soaking time on nutrient bioaccessibility and peptide formation in 30-months-ripened Parmigiano Reggiano cheese. Molecules. 2022;**27**(3):664

[16] Nguyen DD, Johnson SK, Busetti F, Solah VA. Formation and degradation of beta-casomorphins in dairy processing. Critical Reviews in Food Science and Nutrition. 2015;**55**(14):1955-1967

[17] Ponte M, Maniaci G, Di Grigoli A, Gannuscio R, Ashkezary MR, Addis M, et al. Feeding dairy ewes with fresh or dehydrated sulla (*Sulla coronarium* L.) forage. 2. Effects on cheese enrichment in bioactive molecules. Animals. 2022;**12**(18):2462

[18] Rafiq S, Gulzar N, Sameen A, Huma N, Hayat I, Ijaz R. Functional role of bioactive peptides with special reference to cheeses. International Journal of Dairy Technology. 2021;**74**(1):1-16

[19] Tynybayeva I, Zhakupova G, Tultabayeva TC, Nurtayeva A, Sagandyk A, Bekbay S, et al. Isolation and screening of active cultures of lactic acid bacteria for inclusion

in the composition of the starter in the preparation of cheese from whey. Eurasian Journal of Applied Biotechnology. 2022;**4**:75-82

[20] Sgarbi E, Lazzi C, Tabanelli G, Gatti M, Neviani E, Gardini F. Nonstarter lactic acid bacteria volatilomes produced using cheese components. Journal of Dairy Science. 2013;**96**(7):4223-4234

[21] Kurbanova M, Voroshilin R, Kozlova O, Atuchin V. Effect of lactobacteria on bioactive peptides and their sequence identification in mature cheese. Microorganisms. 2022;**10**(10):2068

[22] Lepilkina O, Grigorieva A. Enzymatic proteolysis during the conversion of milk into cheese. Food Systems. 2023;**6**(1):36-45

[23] Helal A, Pierri S, Tagliazucchi D, Solieri L. Effect of fermentation with Streptococcus thermophilus strains on in vitro gastrointestinal digestion of whey protein concentrates. Microorganisms. 2023;**11**(7):1742

[24] Araújo-Rodrigues H, Martins AP, Tavaria FK, Dias J, Santos MT, Alvarenga N, et al. Impact of LAB from Serpa PDO cheese in cheese models: Towards the development of an autochthonous starter culture. Food. 2023;**12**(4):701

[25] Sturova YG, Grishkova AV, Konshin VV. Assessment of the correlation between biochemical processes and product quality in the development of noble mold cheese biotechnology. Proc Univ: Appl chem. Biotechnol. 2022;**12**(43):566-575

[26] Samelis J, Tsanasidou C, Bosnea L, Ntziadima C, Gatzias I, Kakouri A, et al. Pilot-scale production of traditional galotyri PDO cheese from boiled ewes'

milk fermented with the aid of greek indigenous Lactococcus lactis subsp. cremoris starter and Lactiplantibacillus plantarum adjunct strains. Fermentation. 2023;**9**(4):345

[27] Moiseenko KV, Begunova AV, Savinova OS, Glazunova OA, Rozhkova IV, Fedorova TV. Biochemical and genomic characterization of two new strains of *Lacticaseibacillus paracasei* isolated from the traditional corn-based beverage of South Africa, mahewu, and their comparison with strains isolated from kefir grains. Food. 2023;**12**(1):223

[28] Helal A, Nasuti C, Sola L, Sassi G, Tagliazucchi D, Solieri L. Impact of spontaneous fermentation and inoculum with natural whey starter on peptidomic profile and biological activities of cheese whey: A comparative study. Fermentation. 2023;**9**(3):270

[29] Rehman MAU, Ashfaq K, Ashfaq T, Ghaffari MA, Ali N, Kazmi F, et al. The antithrombotic potential of bioactive peptides induced by buffalo milk probiotic cheddar cheese: Potential of bioactive peptides induced by buffalo milk probiotic cheddar cheese. Pakistan Biomedical Journal. 2022;**5**(6):324-328

[30] Pontonio E, Montemurro M, De Gennaro GV, Miceli V, Rizzello CG. Antihypertensive peptides from ultrafiltration and fermentation of the ricotta cheese exhausted whey: Design and characterization of a functional ricotta cheese. Food. 2021;**10**(11):2573

[31] Martini S, Solieri L, Cattivelli A, Pizzamiglio V, Tagliazucchi D. An integrated peptidomics and in silico approach to identify novel antidiabetic peptides in Parmigiano-Reggiano cheese. Biology. 2021;**10**(6):563

[32] Shafique B, Murtaza MA, Hafiz I, Ameer K, Basharat S, Mohamed Ahmed IA. Proteolysis and therapeutic potential of bioactive peptides derived from Cheddar cheese. Food Science & Nutrition. 2023. pp. 1-16

[33] González-Martín MI, Vivar-Quintana AM, Revilla I, Salvador-Esteban J. The determination of fatty acids in cheeses of variable composition (cow, ewe's, and goat) by means of near infrared spectroscopy. Microchemical Journal. 2020;**156**:104854

[34] Phogat S, Dahiya T, Jangra M, Kumari A, Kumar A. Nutritional benefits of fish consumption for humans: A review. International Journal of Environment and Climate Change. 2022;**12**(12):1443-1457

[35] Satir G, Akturk KU, Yavuz M, Koknaroglu H. Effects of adding rumenprotected palm oil in diet on milk fatty acid profile and lipid health indices in Kivircik ewes. Tropical Animal Health and Production. 2023;**55**(3):159

[36] Zong L, Gao R, Guo Z, Shao Z, Wang Y, Eser BE. Characterization and modification of two self-sufficient CYP102 family enzymes from *Bacillus amyloliquefaciens* DSM 7 with distinct regioselectivity towards fatty acid hydroxylation. Biochemical Engineering Journal. 2021;**166**:107871

[37] Bianchi AE, Zortea T, Cazzarotto CJ, Machado G, Pellegrini LG, dos Santos Richards NSP, et al. Addition of palm oil in diet of dairy ewes reduces saturates fatty acid and increases unsaturated fatty acids in milk. Acta Scientiae Veterinariae. 2018;**46**:10-24

[38] Zajác P, Čurlej J, Čapla J, Benešová L, Jakabová S, Partika A, et al. Fatty acid profile of traditional Slovak ewe's and cow's lump cheese. International Journal of Food Properties. 2023;**26**(1):1426-1444

#### *Cheese's Bioactive Peptide Content and Fatty Acids Profile DOI: http://dx.doi.org/10.5772/intechopen.112712*

[39] Ellis KA, Innocent GT, Mihm M, Cripps P, McLean WG, Howard CV, et al. Dairy cow cleanliness and milk quality on organic and conventional farms in the UK. The Journal of Dairy Research. 2007;**74**(3):302-310

[40] Stergiadis S, Leifert C, Seal C, Eyre M, Larsen MK, Slots T, et al. A 2-year study on milk quality from three pasture-based dairy systems of contrasting production intensities in Wales. The Journal of Agricultural Science. 2015;**153**(4):708-731

[41] Ward A, Judge J, Delahay R. Farm husbandry and badger behaviour: Opportunities to manage badger to cattle transmission of *Mycobacterium bovis*? Preventive Veterinary Medicine. 2010;**93**(1):2-10

[42] Bobbo T, Ruegg P, Stocco G, Fiore E, Gianesella M, Morgante M, et al. Associations between pathogenspecific cases of subclinical mastitis and milk yield, quality, protein composition, and cheese-making traits in dairy cows. Journal of Dairy Science. 2017;**100**(6):4868-4883

[43] Butler G, Nielsen JH, Slots T, Seal C, Eyre MD, Sanderson R, et al. Fatty acid and fat-soluble antioxidant concentrations in milk from high-and low-input conventional and organic systems: Seasonal variation. Journal of the Science of Food and Agriculture. 2008;**88**(8):1431-1441

[44] Stergiadis S, Berlitz CB, Hunt B, Garg S, Givens DI, Kliem KE. An update to the fatty acid profiles of bovine retail milk in the United Kingdom: Implications for nutrition in different age and gender groups. Food Chemistry. 2019;**276**:218-230

[45] Ivanova M. Conjugated linoleic acid-enriched dairy products: A review. Journal of Microbiology, Biotechnology and Food Sciences. 2021;**10**(5):e3609-e

[46] Newton EE, Lamminen M, Ray P, Mackenzie AM, Reynolds CK, Lee MR, et al. Macromineral and trace element concentrations in milk from finnish Ayrshire cows fed microalgae (*Spirulina platensis*) and rapeseed (*Brassica napus*). Journal of Dairy Science. 2022;**105**(11):8866-8878

[47] Ellis KA, Innocent G, Grove-White D, Cripps P, McLean W, Howard C, et al. Comparing the fatty acid composition of organic and conventional milk. Journal of Dairy Science. 2006;**89**(6):1938-1950

[48] Ormston S, Qin N, Faludi G, Pitt J, Gordon AW, Theodoridou K, et al. Implications of organic dairy management on herd performance and milk fatty acid profiles and interactions with season. Food. 2023;**12**(8):1589

[49] Dehareng F, Delfosse C, Froidmont E, Soyeurt H, Martin C, Gengler N, et al. Potential use of milk mid-infrared spectra to predict individual methane emission of dairy cows. Animal. 2012;**6**(10):1694-1701

[50] Khalesi M, Cermeño M, FitzGerald RJ. Contribution of whey protein denaturation to the in vitro digestibility, biological activity and peptide profile of milk protein concentrate. Journal of Functional Foods. 2023;**104**:105543

[51] Yildirim SC, Ates F. Antimicrobial edible cellulose-based (CB) films and coatings for enhancing microbial safety of white cheese during storage. Emirates Journal of Food and Agriculture. 2022;**34**(12):1061-1071

[52] Mirzapour-Kouhdasht A, McClements DJ, Taghizadeh MS, Niazi A, Garcia-Vaquero M. Strategies for oral delivery of bioactive peptides with focus on debittering and masking. npj Science of Food. 2023;**7**(1):22

[53] Pop OL, Kerezsi AD, Ciont C. A comprehensive review of *Moringa oleifera* bioactive compounds—Cytotoxicity evaluation and their encapsulation. Food. 2022;**11**(23):3787

[54] Inácio RS, Rodríguez-Alcalá LM, Pimentel LL, Saraiva JA, Gomes AM. Evolution of qualitative and quantitative lipid profiles of high-pressureprocessed Serra da Estrela cheese throughout storage. Applied Sciences. 2023;**13**(10):5927
