Camel Milk's Cheese and Paneer Cheese

#### **Chapter 5**

## Recent Trends on Camel Milk Cheese Processing: Nutritional and Health Value

*Zeineb Jrad, Olfa Oussaeif and Halima El-Hatmi*

#### **Abstract**

The capacity of dairy components to prevent chronic diseases has piqued researchers' interest in the role they play in the creation of functional meals. In this regard, the demand for camel milk has increased dramatically due to its outstanding therapeutic properties and health-promoting effects. Ever since ancient times, camel milk has only ever used unprocessed for the consumption of the nomads and their own families. The limited use of camel milk is due to its manufacturing difficulties. For a long time, cheese-making from camel milk was considered a challenge, due to its unique composition. However, due to the development of processes, and enzymatic and microbial technologies, the dairy sector is now able to offer consumers camel cheese with improved functionality and nutritional advantages. The current chapter highlights the recent processing opportunities regarding the cheese-making from camel milk and summarizes existing knowledge on the nutritional value of camel milk cheese.

**Keywords:** camel milk, cheese, nutritional value, dairy processing, biological activities

#### **1. Introduction**

Currently, great importance is given to nutrition as a vector of health. Milk is one of the food substances considered very nourishing and necessary for the growth of young children. Over the years, a particular attention has been paid to milk from nonbovine species such as goat, ewe, mare, and camel. The camel milk sector is gaining importance due to the particular composition of this milk and its potential value in improving the consumer health. Indeed, some people start consuming camel's milk because they think that camel's milk can cure various diseases, such as jaundice, diabetes, ulcers, autism, asthma, allergies, and cancer [1]. Several studies have provided an opportunity to understand the potential benefits of camel milk through the production of bioactive peptides. Thus, numerous such peptides are identified such as angiotensin-converting enzyme inhibitors, antimicrobial, anti-inflammatory, anti-obesity, antioxidant as well as antidiabetic and anti-proliferative peptides [2–5]. In recent years, the research on camel milk transformation has gradually increased. Much of the work in this field has focused on making yogurt, butter, ice cream, and

cheese from camel milk. However, one of the major issues confronting the camel milk cheese-making is the low yield, the long coagulation time, the watery consistency, and the fragile and poor structure, affecting both product performance and customer sensory perception [6]. This behavior is probably due to the large size of casein micelles [7], the low amount of κ-casein to β-casein ratio and its specific enzyme cleavage site, and the lack of β-lactoglobulin in camel milk [8]. The small size of camel milk fat globule could be another cause of poor gel structuring in camel milk [9].

Many attempts have been made to solve this problem including the use of enzymes as coagulants and/or the addition of acidification of the milk with different starter cultures or acids [10–15]. Some researchers mixed camel milk with other milk in order to enhance the yield and organoleptic properties of camel cheese [16–18], whereas others used plant protease [19–22] or microbial transglutaminase [22–24]. Additionally, it could be possible to overcome the associated difficulty regarding camel milk cheese-making through the application of new technologies [8, 25] or optimization of processing conditions [7, 26–28].

In the current chapter are highlighted advanced approaches used in camel cheesemaking process steps. In addition, selected camel cheese varieties' specific characteristics and their typical nutritional value and functionalities are also described.

#### **2. Constraints associated with camel's milk cheese-manufacturing**

Cheese is one of the most consumed dairy products worldwide due to its tremendous nutritional benefits such as richness of proteins, fatty acids, calcium, vitamins A and B12, and bioactive peptides coming from milk fermentation with lactic acid bacteria [29]. Cheese is usually produced from casein coagulation and precipitation following acidifying or/and fermentation.

The main factors responsible for crud formation are listed as follows:


Moreover, these different components have particularities in camel milk. It is found that whey proteins in camel milk account for 20–25% [30]. As in human milk, the major and most allergenic protein of bovine milk, β-lactoglobulin (β-LG), is not detected in camel milk, but it is the α-lactalbumin (α-LA) which is the most abundant protein [31, 32]. The absence of β-LG in camel milk presents itself as an obstacle in the camel cheese-making. In fact, the implication of β-LG on milk transformation is important mainly through its heat-induced association with κ-CN.

Since heat treatment is necessary for eliminating harmful bacteria, extending the shelf life of milk, and guaranteeing its safety for human consumption, it is a crucial component of the dairy processing sector [33]. It is known that the manufacture of

#### *Recent Trends on Camel Milk Cheese Processing: Nutritional and Health Value DOI: http://dx.doi.org/10.5772/intechopen.114032*

cheese is always preceded by a step of milk heat treatment (pasteurization). Differing from cow milk, camel milk is less stable and more sensitive to various heat treatments and this instability provides a crud with a weak structure [34]. It is important to note that casein distribution and micelle size are limiting factors in cheese coagulation. The primary structure of the four caseins was elucidated by Kappeller et al. [35] deducing that camel caseins are less phosphorylated and less rich in micellar calcium phosphate than their bovine counterparts. The camel casein micelles are also larger in diameter than bovine milk casein, this character associated with reduced surface area provided a long coagulation time and weak cheese coagulum [36]. Camel's milk caseins, on the other hand, are richer in proline residues (particularly β-casein), residues known by their stereo-chemical rigidity, thus explaining the destabilization of the secondary structures of these proteins in a more pronounced way than occurs in bovine caseins. Camel κ-casein represents 3.5% of total camel caseins and it contains two phosphate residues present in two positions: Ser141 and Ser159. Only κ-casein is a glycoprotein with amphiphilic properties. Camel and bovine κ-casein do not have the same affinity for camel and bovine rennet calf [35].

The cleavage site of camel κ-casein by chymosin is different from that of bovine κ-casein, the hydrolysis taking place at the Phe97-Ile98 bond splitting a macro-peptide of 65 amino acids. A classification into two groups of κ-caseins of different species has been proposed by Nakhasi et al. [37]. These two groups differ in the site of cleavage by chymosin: Phe-Met bond for group I (ewe, buffalo, goat, and cow) and Phe-Ile or Phe-Leu for group II (Camel, woman, rat, mouse, and sow). This difference probably reflects differences in the ability of ruminant and non-ruminant milk to coagulate.

The protein κ-casein is considered the constituent limiting the growth of submicelles as well as the size of the micelles [35]. It is also the stabilizing factor of the micelle thanks to the hydrophilic C-terminal groups of this protein which are responsible for the steric repulsive forces, which oppose the flocculation of the micelles [38]. For these reasons, κ-casein is most likely localized to the periphery of the casein micelle.

On the other hand, the high amount of β-casein seems to have an important implication in the softer aspect of camel cheese compared to bovine one. It is the major protein in camel milk and represents more than 65% of total caseins. As compared to the other milk proteins, CN-β is more hydrophobic and exhibits more chaperone-like behaviors, which prevents protein aggregation [35]. Moreover, CN-β with its amphiphilic nature rises for non-polar residues to adsorb at hydrophobic surfaces, resulting in good emulsifying properties that are responsible for the smoothness of cheese [39]. Apart from this, milk fat also plays an important role in cheese quality and yield. Camel milk fat globules, surrounded by thick membranes, are small in size ranging from 1 to 9 μm in diameter according to Mehaïa et al. [40]. Camel milk fat is also distinguished by high levels of phospholipids [41], which are good emulsifiers, that give camel milk cheese its soft texture and great water retention.

#### **3. Camel milk cheese historical background and recent insights into cheese-making**

In the past, people used the process of creating cheese to preserve milk. The first cheese was made in the United States. There were more than 45 million kg of cheese manufactured in the United States in 1849. Raw milk was used to make the majority of the cheese. Despite being a novel technology, pasteurizing milk for making cheese was not widely practiced in 1914. The most popular cheeses today are Parmesan and

Gorgonzola from Italy, Emmental from Switzerland, Roquefort and Camembert from France, and Edam from Holland. Since then, cheese has gained enormous popularity throughout the world. Research on cheese from minor milk species has increased in the last 30 years, especially for camel milk. This interest was initially influenced by the difficulties of camel's milk coagulation. Furthermore, its technologies that are currently being applied for the cow's cheese-making are not successfully applicable to turn camel milk into cheese. Earlier attempts at creating camel cheese involved combining camel milk with goat and sheep milk and the use of bovine rennet, but the obtained coagulum was extremely soft and crumbly [42, 43]. After that, the difficulty of camel milk coagulation and the low cheese's yield has been confirmed by the author's observations [44, 45] who are into the idea of overdosing the rennet concentrations about four times more than that used for cow milk. The used rennet was that of the calf combined with a coagulant preparation derived from a mold conventionally used in the dairy industry named *Rhizomucor miehei*. Additionally, they demonstrated how the camel milk may be supplemented with calcium chloride or calcium phosphate to somewhat sidestep this challenge. In fact, salts modify the ionic environment of the casein micelle and bring about a lowering of the pH value of the milk, which promotes the activity of the rennet and the process of aggregation of the casein leading to the coagulation. Unlike, the obtained cheese turned out to be fragile and crumbly, the cheese's yield was low with high loss in fat-which led to not creamy product.

More recently, many researchers used Camifloc—a specific rennet used to coagulate camel milk—as a coagulant [28, 46]. Later, some authors used Camifloc to coagulate camel milk either by adding sheep's milk at 50 and 75% levels [47] or by varying the levels of added salts [28]. In concert, these results demonstrate a reduction in coagulation time and an improvement of the texture of cheese as well as sensory appreciation. Other alternative has been considered such as introducing the camel chymosin synthesis gene into *Aspergillus niger* to obtain a specific recombinant enzyme for camel milk coagulation named Chymax-M1000® produced by Ch. Hansen©. Hence, the camel cheese qualities were enhanced [10, 26]. Then, several studies have demonstrated that the association of camel chymosin with starter cultures improves the camel cheese yields. Different starters were added like thermophilc, mesophilic or blended strains, and yogurt starters [10, 19, 21, 48, 49]. Walle et al. [50] reported that cooking camel milk coagulated with camel chymosin combined with starter cultures improved the hardness of cheese.

The production issues related to camel cheese were resolved by mixing the milk of other bovines such as cow, buffalo, sheep, and goat milk in order to boost the casein concentration in camel milk. According to the research of Shahein et al. [17], the 30% (w/w) addition of buffalo milk to camel milk improved the rennet's ability to coagulate, increased curd yield, improved curd hardness, reduced weight loss, and improved the sensory and microbiological quality of the finished product [17]. However, Saadi et al. [18] produced soft cheese by adding sheep's milk to camel milk, and discovered a significant improvement in the yield and cheese quality.

Moreover, processing conditions could affect greatly the quality and the nutritional value of produced cheese. For instance, raising the total solid content in camel milk using the ultrafiltration process was found to increase the cheese yield, firmness, and nutritional value due to the end product's higher protein and fat, in comparison with conventional processing [8, 51]. Furthermore, the addition of *Allium roseum* powder to UF camel milk produces a camel cheese with higher anti-oxidant activities [8]. Likewise, it has been documented that cheese hardness is significantly influenced

#### *Recent Trends on Camel Milk Cheese Processing: Nutritional and Health Value DOI: http://dx.doi.org/10.5772/intechopen.114032*

by the milk pasteurization temperature, high-pressure treatment, and pre-acidification. Mbye et al. [25] showed that camel milk pasteurization at temperatures not exceeding 65°C for 30 min or high-pressure processing at 350 MHz for 5 min at 4°C are more effective in providing cheeses from camel milk with a semi-hard texture.

In fact, the most notable modifications during cheese-making are observed after high-pressure processing or homogenization resulting in the reduction of the size of the native milk fat globule. In addition to that, high-pressure processing alters the conformational shape of casein micelles by reducing electrostatic and hydrophobic interactions, which causes micellar fragments to disaggregate and improves milk's physico-chemical and technological applications. Casein micelles are broken apart, increasing the surface area and hastening the rennet coagulation process. Consequently, this treatment can lead to the formation of new clusters of fat and protein, providing the opportunity for many different cheese textures. Some research has come out and has been studied the effect of pre-acidification technology on camel cheese aspect. The pre-acidification of the milk before adding the enzyme can be done by adding directly acids or by using bacteria that can metabolite lactic acid. In both cases, the presence of acid in the milk attributes to the decrease of its pH value and decreases the coagulation time of camel milk [10–15].

Other trials have been tested by various authors in order to improve camel milk cheese's textural and sensorial qualities. These include the use of rennet substitutes of animal origin such as bovine pepsins and pepsins extracted from poultry proventriculi such as chicken and duck [18, 52]. Recently, a great deal of research has been undertaken in order to find effective and competitive coagulants using plant extracts such as ginger (*Zingiber officinale*) [53], *Moringa oleifera* L. [20], Withania (*Withania coagulans*) [21], and nettle (*Urtica dioica*) leaves [19]. The results of these studies showed that the ability of partially purified plant extracts to coagulate camel milk and form a firm curd. It has also been demonstrated that camel milk cheese produced using a combination of withania extract and camel chymosin exhibited a higher quality when compared to chymosin alone [21]. At the same time, other studies have been devoted to improving camel milk coagulation using microbial transglutaminase (MTGase). Abou-Soliman et al. [23] reported that adding MTGase to camel milk at a concentration of 80 U after 20 min of renneting is recommended to improve the yield, textural, and some sensory properties of soft cheese made from camel milk. Alia et al. [22] found that MTGase and the *Cynara cardunculus* L. flower extracts showed excellent coagulating properties and immense potential as coagulants for cheese production using camel milk.

#### **4. Camel cheese nutritional value and bioactive components**

#### **4.1 Substances composition of camel cheese**

The variation of composition observed in the camel cheese may be due to the original milk composition and cheese-making processing circumstances. The total solids components, including protein and fat, progressively concentrate into the cheese curd depending on how the cheese is prepared and how the whey is drained. Additionally, the kind, amount of ash, and salt addition can all affect the minerals in the cheese during the cheese-making process. In addition to its impact on milk clotting, the acidification process is essential for the removal of colloidal minerals from casein micelles, coagulant retention in the curd, coagulum strength, and cheese yield. The chemical composition of the different camel cheeses is detailed in **Table 1**.


#### **Table 1.**

*Composition and yield of different types of camel milk cheese (%).*

#### **4.2 Proteins**

Protein is one of the main nutritional substances in camel milk cheese. The protein content in camel milk directly affects the nutritional value of camel dairy products. The protein content in camel milk cheese is affected by several factors, such as the type of starter culture used for cheese-making, meaning that a camel cheese made with

*Recent Trends on Camel Milk Cheese Processing: Nutritional and Health Value DOI: http://dx.doi.org/10.5772/intechopen.114032*

thermophilic (STI-12) and blended (RST-743 and XPL-2) cultures had a significantly higher protein value [15]. The variation of protein content into the categories of camel cheese obtained might be attributed to the processing condition of cheese manufacture. The ultrafiltration process and cheese fortification enhanced significantly the protein content of soft camel cheese [8]. Moreover, the protein content into the cheese curd depends on how the whey is drained. Besides mixing camel milk with other dairy animal milk has a substantial effect on the protein amount in camel cheese [58].

#### **4.3 Fat**

Fat is an important factor that may be responsible for cheese quality. Fat contents are progressively concentrated into the cheese curd according to cheese processing and the method used to drain the whey. Castillo [63] reported that the rheological and micro-structural properties of gels' strength and the higher curd loss from the cheese vat resulted in excess whey fat loss. A huge variation of fat content in camel mozzarella cheese blends of bovine milk and 30% of camel milk [58]. A significant difference in fat cheese was observed in soft white cheese with different starter cultures [15], whereas the variation of percentages of salt to make Domiati-type camel cheese has no effect on fat content [12]. El-Hatmi et al. [8] reported a low content in soft camel cheese due to a loss of fat in permeate during the ultrafiltration process.

#### **4.4 Total solids**

According to the studies focused on camel cheese characterization, there is a variation in total solids observed, and this variation might be attributed to the original milk composition on protein and fat, the processing condition of cheese manufacturing and the method used in why draining. In fact, total solids are concentrated into the cheese curd. The acidifying process of milk during cheese-making, is a determining factor in the dry matter content of cheese, and this is due to its important role in the removal of colloidal minerals from casein micelles, the retention of coagulant in the curd, syneresis of the gel, coagulum strength, and cheese yield [64, 65].

#### **4.5 Biological properties**

The bioactive peptides derived from camel milk proteins and products, particularly fermented milk, have received much attention during the last decade. However, limited studies have been done with camel cheese as a source of bioactive peptides. There are mainly two approaches used to produce bioactive peptides from camel milk, that is, bacterial fermentation and enzymatic hydrolysis. Identified camel peptides of single and/or multiple functions have been reported as follows:

#### • *Anti-oxidant peptides:*

Several anti-oxidant peptides have been obtained from the action of digestive enzymes or lactic acid bacteria proteinase on camel milk casein. These peptides have been identified as fragments of a camel β-CN, α-CN, GlyCAM-1, and PGRP-1 [2, 66, 67]. To our knowledge, antioxidant peptides of camel cheese have never been identified, but the anti-oxidant activity of camel milk cheese was well documented. El-Hatmi et al. [8] reported that UF camel milk cheese exhibited antioxidant activity and this power was improved after cheese fortification with *Allium roseum* powder [8] or quinoa flour [68].

Whereas Abou Soliman et al. [23] showed that the cross-linking between camel-milk proteins caused by MTGase negatively influenced the antioxidant activity of cheese.

#### • *Antimicrobial peptides:*

The presence of antimicrobial activity in the camel whey protein digests has also been reported [2]. Jrad et al. [69] identified cationic peptides from the peptic digests of camel lactoferrin (LF). They found these peptides to have antimicrobial activities against *Listeria innocua*. Digestion of camel LF with pepsin resulted in an antimicrobial peptide homologous to cow lactoferrampin B. The camel lactoferrampin represents LF fragment f 271–284 with the sequence LVKAQEKFGRGKPS [69]. In addition, other antibacterial peptide derived from a camel β-CN was identified and presented high homology with casesidin bovine peptide [70]. Low molecularidentified antimicrobial peptides were generated from fermented camel milk with *Lactobacillus plantarum* [71].

#### • *ACE-inhibitory peptides:*

The ACE-inhibitory peptides found in hydrolyzed camel proteins and in products of camel milk have received much attention. The ACE-inhibitory peptides isolated from the digests of camel milk casein with potential activity are dipeptides ("AI," "IY," "VY," "LY," "TF") and tri-peptides ("IPP," "LHP") [72]. The angiotensin-converting enzyme inhibition of camel cheese was also investigated, but there is no information about derived peptides [73].

#### • *Anti-diabetic peptides:*

Camel milk constitutes a center of interest for scientists due to its known beneficial impact on diabetes. Identification of camel milk-derived peptides and their structure-activity relationship study and characterization in the context of molecular markers related to diabetes are studied. The main targeted enzymes for their inhibition by camel milk proteins/peptides are carbohydrate digestive enzymes, specifically intestinal α-glucosidase and pancreatic α-amylase, and the dipeptidyl peptidase IV (DPP-IV), an enzyme that breaks down major incretin hormones that stimulate the release of insulin in response to glucose [3, 74]. It is now obvious that identified antidiabetic peptides that target the key molecular pathways involved in overall glucose homeostasis liberated from both camel milk whey and casein [75]. Moreover, camel cheese exhibited α-amylase and α-glucosidase inhibition activity [73].

Extensive other *in vitro* studies have found that camel milk and its derived products possess anti-obesity, anti-biofilm, anti-cancer, anti-inflammatory, antihemolytic, and anti-hyperpigmentation activities. This provides potential for the development of functional products using camel milk.

#### **5. Conclusions**

Worldwide, camel milk and its derivative products production and consumption have increased due to its medicinal and health-promoting potential, which makes it the best choice as a substitute of cow's milk. However, the processing methodology of camel milk into dairy products is facing several difficulties. In fact, making cheese *Recent Trends on Camel Milk Cheese Processing: Nutritional and Health Value DOI: http://dx.doi.org/10.5772/intechopen.114032*

from camel milk using the same conventional methods used for cheese manufacture from cow's milk is challenging and occasionally impossible because of a number of issues, such as prolonged coagulation times, weak curd formation, and ultimately lower cheese yield. Therefore, with current advancements in dairy technology, the processing of camel milk into cheese become now possible. Camel milk cheeses made with camel chymosin, with starting cultures and through technological processing, have significantly improved in some cheese-making qualities. Moreover, it was discovered that clotting camel milk with plant protease and chymosin is a successful method to make cheese from camel milk. Additionally, camel milk has significant enhancement in various features of cheeses when combined with other milk species. Some studies have focused on the functional characteristics and nutritional quality of camel milk cheeses, but additional clinical studies are required to confirm the therapeutic effects of these functionalities in the human body.

#### **Conflict of interest**

The authors declare that they have no conflict of interest.

#### **Author details**

Zeineb Jrad1 \*, Olfa Oussaeif<sup>2</sup> and Halima El-Hatmi1

1 Department of Agrifood, Higher Institute of Applied Biology of Medenine, University of Gabes, Tunisia

2 Livestock and Wildlife Laboratory, Institute for Arid Regions of Medenine, Tunisia

\*Address all correspondence to: jradzeineb@yahoo.fr

© 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.

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[58] Abdalla A, Abu-Jdayil B, Alsereidi H, Hamed F, Kamal-Eldin A, Huppertz T, et al. Low-moisture part-skim mozzarella cheese made from blends of camel and bovine milk: Gross composition, proteolysis, functionality, microstructure, and rheological properties. Journal of Dairy Science. 2022;**105**(11):8734-8749

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[75] Ayoub MA, Yap PG, Mudgil P, Khan FB, Anwar I, Muhammad K, et al. Camel milk-derived bioactive peptides and diabetes: Molecular view and perspectives. Journal of Dairy Science. 2023 [In press]

#### **Chapter 6**

## Indian Cheese Revolution: *Withania coagulans* in Dairy Industry

*Mayur Ram, Bimal S. Desai and Sumankumar S. Jha*

#### **Abstract**

Commonly called as Indian Cheese Maker, Paneer dodi, Paneer phool and Vegetable rennet, *Withania coagualans* belongs to family Solanaceae and widely used in Indian System of Medicines due to its anti-diabetic, anti-microbial and immune modulator properties. The specific epithet coagulans reveals its coagulating properties and hence used in Punjab and parts of Northern India in cheese and paneer industries. The plant is rich in enzyme Withanin which is responsible for the coagulating properties. Many herbal prescriptions like Liv 52, (Liver Health Support Supplement) which is an Ayurvedic poly herbal formulation consists of extracts of both *Withania somnifera* and *Withania coagulans.* Commercial cultivation of this plant is in its initial phase in parts of Punjab, Haryana and also in neighboring countries as it has wide distribution extending up to South Asia. Plant is also rich in Withanolide contents and can be a future prospect for South Gujarat region, as coagulating agent for dairy industries and rennet enzyme production. Seeds are available in local markets of Surat and Navsari, routinely used for controlling diabetes. The chapter aims at the possibilities of cultivating this plant in South Gujarat conditions in India, since the other species *Withania somnifera* is also available and has naturalized in AES Zone III of South Gujarat.

**Keywords:** rennet, Withanin, withanolides, poly herbal, anti-diabetic

#### **1. Introduction**

South Asia is home to many rich Traditional Systems of Medicine (TSM) including Ayurvedic, Unani, Siddha and Tibetan systems, which have been helpful in sustaining healthy life of tens of millions of people for centuries. India possesses abundant reserves of medicinal and aromatic plants (MAPs) across a vast territory characterized by diverse environmental conditions. The strategic geographical location, unique geomorphology, the existence of ancient flora from geological eras, and the harmonious interplay between biotic and non-biotic factors have collectively contributed to India being recognized as a region of remarkable plant diversity and endemism. These factors directly influence the wide range of medicinal and aromatic plant species found within the country. Himalayan sage scholars of TSM have said "*Nanaushadhi Bhootam Jagat Kinchit*" i.e., there is no plant in the world, which does not have

medicinal properties.' These venerable scholars possessed extensive knowledge about the medicinal properties of numerous plant species, with estimates suggesting their understanding extended to hundreds of plants. It is not an overstatement to assert that the utilization of plants for enhancing human health dates back to the very origins of human existence [1].

The genus Withania, belonging to the Solanaceae family, holds a prominent position in the Indian Ayurvedic system of medicine due to its remarkable pharmaceutical and nutraceutical properties. Out of the 26 identified species within the genus Withania, only two species, namely *Withania somnifera* (L.) Dunal and *Withania coagulans* (Stocks) Dunal, have garnered significant economic importance [2]. Withania has been widely utilized in traditional folk medicine for treating a wide range of ailments. Additionally, one particular species, *W. coagulans*, is commonly referred to as the Indian cheese maker or Vegetable rennet. This species has been traditionally employed in various regions of India for the preparation of rennet ferment used in cheese production from vegetables. Different parts of this plant have been reported to possess a variety of biological activities [3].

Since decades ago, fruits of this plant were used widely in the production of traditional cheese from raw cow's milk. It is believed that the milk coagulation ability corresponds to the presence of an enzyme in the berries of the plant as it is related to the pulp and husk of the berry. Cottage and Cheddar cheese can be made using fruit extracts of *W. coagulans* as a good alternative to animal rennet, though the cheese produced with the described extract has a notable bitter flavor that can be reduced by increasing the ripening time [4].

#### **1.1 Indian cheese: a wealth of health benefits**

Indian cheese, particularly varieties like paneer has been a staple in Indian cuisine for centuries. Beyond its culinary appeal, Indian cheese offers a plethora of health benefits, making it a popular choice not just for its taste, but for its nutritional value as well.


for nerve health and red blood cell production, while zinc supports the immune system and promotes wound healing.


### **2. Distribution**

*W. coagulans* is distributed in the East of the Mediterranean region extending to South Asia i.e., Iran, Afghanistan, Pakistan (Sind and Baluchistan), Nepal and India, up to 1700 m. In India, it is found in (North-West India) Himachal Pradesh, Punjab, Uttarakhand and Rajasthan. In Rajasthan it is sporadically distributed in Barmer, Jaisalmer and Jodhpur districts of Western Rajasthan desert and it is not common, categorized as "vulnerable species" by Pandey *et al.* [5]. It's important to note that *Withania coagulans* has also been introduced and cultivated in other parts of the world, including certain regions of the United States, Australia, and Europe. However, its natural distribution is primarily cantered in South Asia. Overall, *Withania coagulans* is adapted to grow in arid and semi-arid environments and is commonly found in the regions mentioned above.

### **3. Cultivation**

*W. coagulans*, commonly known as Indian Rennet or "Paneer Doda," is primarily cultivated in specific regions where it is native or adapted to grow. The main cultivation areas for *Withania coagulans* include:


Outside of its native range, *W. coagulans* has also been introduced and cultivated in some other countries, such as the United States, Australia, and parts of Europe. However, the cultivation in these areas might be limited and less widespread compared to its native regions. It's worth noting that successful cultivation of *W. coagulans* requires specific growing conditions, including well-drained soil, warm temperatures, and a semi-arid climate. The plant is well-adapted to arid and dry regions, and its cultivation is typically focused in areas that provide the necessary environmental factors for its growth.

*W. coagulans* cultivation is primarily concentrated in India. India is one of the main countries where *Withania coagulans* is cultivated due to its suitability for the plant's growth requirements and its traditional use in Ayurvedic medicine. In India, the cultivation of *W. coagulans* is particularly prominent in regions with arid and semiarid climates. States like Rajasthan, Gujarat, Punjab, and Haryana are known for their significant cultivation of *W. coagulans*. These areas provide the necessary conditions,

**Figure 1.** *Morphology of Withania coagulans.*

*Indian Cheese Revolution:* Withania coagulans *in Dairy Industry DOI: http://dx.doi.org/10.5772/intechopen.113815*


#### **Table 1.**

*Taxonomical classification.*


#### **Table 2.**

*Synonyms.*

including dry and warm climates, well-drained soil, and limited rainfall, which are favorable for the growth of *W. coagulans*.

Farmers in these regions have been cultivating *W. coagulans* for generations and have developed knowledge and expertise in its cultivation practices. The plant is typically grown as a cash crop by farmers who recognize its value in traditional medicine and Paneer Industries demand in the market. Overall, India is a major contributor to the cultivation and production of *W. coagulans*, meeting both domestic demand and supplying it to various other countries for its medicinal and therapeutic applications (**Figure 1**, **Tables 1** and **2**).

#### **4. Botanical description**

*W. coagulans* is a sturdy gray undershrub, reaching a height of 60–120 cm. Its flowers from November to April, while the berries ripen from January to May. The natural regeneration is from the seed. The flowers dioceous, in auxiliary clusters; pedicils 0.6 mm long, Deflexed, slender. The calyx of *W. coagulans* is campanulate, measuring 6 mm in length and covered with a delicate stellate gray tomentum. Its teeth are triangular, approximately 2.5 mm long. The corolla of *W. coagulans* measures 8 mm in length and is covered with a stellate mealy texture on the outside. It is divided approximately one-third of the way down, and its lobes are ovate-oblong with a sub-acute shape. Male flowers stamens about level with the top of the corolla -tube; filament 2 mm long, glabrous; anthers 3–4 mm long. Ovary ovoid, without style or stigma. Female flowers stamens scarcely reaching 1/2 way up the corolla-tube; filaments about 0.85 mm long; anther smaller than in the male flowers, sterile. Ovary

ovoid, style glabrous; stigma mushroom-shaped, 2-lamellate. Berry 6–8 mm globose, smooth, closely girt by the enlarged membranous calyx, which is scurfy -pubescent outside. Seeds 2.5–3.0 mm diameter, somewhat ear shaped, glabrous [7, 8].

#### **5. Phytochemistry**

Different chemical components such as alkaloids, hormones, tannins, saponins, carbohydrates, protein, amino acids and organic acid are shown by aqueous and methanolic extracts from the *W. coagulans*. Seeds have 17.8% free sugars, maltose, fatty oil, D-galactose & D-arabinose. The fruit contains a milk-coagulating enzyme, two esterases, free amino acids, essential oil and fatty oil and alkaloids, triacontaine hydrocarbon, dihydrostigmasterol sterol. Proline, hydroxyproline, valine, tyrosine, aspartic acid, glycine, asparagine, cysteine, and glutamic acid are amino acids. Alkaloidal fractions were isolated from the fruit's alcoholic extract. Leaves contain four withanolides-called steroidal lactones, Withaferin-A, 5, 20α(R)-dihydroxy-6α,7αepoxy-1- oxo-(5α)-with a-2,24-dienolide and two minor withanolide, of which one is probably 5α, 17α-dihydroxy-1- oxo-6, 7α-epoxy-22R-witha-2,24- dienolide [9].

The term "withanolide" is a structural designation coined by combining "withan" from the genus Withania with "olide," which is the chemical term for a lactone. Major bioactive phytoconstituents isolated from *W. coagulans* are lactone steroids called withanolides. A new group of steroidal lactones called withanolides has been recently isolated from different species of the solanaceae family, mainly *Withania Somnifera*. The whole plant of *W. coagulans contains* various withanolides, including coagulin F, coagulanolide, withacoagulin, and coagulin G. Additionally, the roots, leaves, and fruits of this plant have been reported to contain four, two, and two withanolides respectively, which serve as important biogenetic precursors of withanolides, withanolides shows antitumurous, anti-inflammatory, antibacterial, immunosuppressive activities [10]. A new withanolide, with a unique chemical structure similar to the aglycones of the cardiac glycosides, was isolated from the fruits of *W. coagulans*, and was screened for cardiovascular effects. The diverse therapeutic applications of withanolides found in *W. coagulans* have garnered significant interest in the scientific community. A new Withanolide isolated from *W. coagulans* have been found to be active against several potentially pathogenic fungi. Withanolides have been reported to possess both immunostimulating and immunosuppressive effects in different studies. Withanolides have been reported to have effect on haemopoietic system and bone marrow. Glycowithanolides have been found to have effects on CNS [11].

Withanolides, which constitute a major component of *W. coagulans*, can be classified chemically into the following groups [6].


### **6. Medicinal use of different parts of** *Withania coagulans*

Different parts of *W. coagulans*, including the leaves, roots, seeds, and fruit, have been traditionally used for their medicinal properties. Here are some of the medicinal uses associated with each part [12].


### **7. Pharmacological properties**

The plant's berries are used to coagulate milk. It has always played a significant role in the Ayurvedic, Unani, and traditional Indian medical systems. Numerous studies have shown that the isolated withanolides from *W. coagulans* have interesting biological properties. The plant's sweet fruits are also said to have sedative, emetic, alterative, and diuretic properties. They are helpful for liver complaints that are persistent. They have occasionally been employed as blood purifiers. Additionally, they are used to treat other intestinal infections, flatulent colic, and dyspepsia. These are used to treat asthma, biliary conditions, and stuttering Maurya [11].

• *Anti-inflammatory*: *W. coagulans* exhibits anti-inflammatory properties, which can help reduce inflammation in the body. These effects are attributed to the presence of bioactive compounds like withanolides, flavonoids, and other phytochemicals.


#### **8. Significance of** *W. coagulans* **in dairy industries**

The primary step in cheese production is milk coagulation. For thousands of years, coagulating enzymes, which are preparations of proteins that break down other proteins, have been utilized in the process of making cheese. Interestingly, this practice of using enzymes in cheesemaking is believed to be one of the earliest known applications of enzymes. The first evidence of cheesemaking can be traced back to cave paintings dating around 5000 BC [16]. Over time, scientists have developed alternatives to animal rennet, such as rennet substitutes produced by microorganisms and genetically engineered microorganisms. However, there is a growing interest in vegetable coagulants, which are milk-clotting enzymes extracted from plants. Cheeses made with vegetable coagulant can be found mainly in Mediterranean, West African, and southern European countries. Spain and Portugal have the largest variety and production of cheeses. *Withania coagulans*, commonly known as Indian Rennet or "Paneer Doda," has traditional uses in the dairy industry. It is used as a natural coagulant or rennet substitute in the process of cheese and paneer (Indian cottage cheese) production.

In traditional cheese making, rennet is commonly used as a coagulant to separate milk into curds and whey. However, *W. coagulans* offers an alternative source of coagulant enzymes that can achieve similar effects. The ripe fruit of *W. coagulans* contains enzymes known as milk-clotting proteases or chymosin-like proteases. These

#### *Indian Cheese Revolution:* Withania coagulans *in Dairy Industry DOI: http://dx.doi.org/10.5772/intechopen.113815*

enzymes have the ability to coagulate milk proteins, resulting in the formation of curds [17]. This property makes *W. coagulans* a natural substitute for animal-based rennet in cheese making, particularly in vegetarian or vegan cheese production. In the dairy industry, *W. coagulans* is sometimes used to produce a variety of cheeses, including traditional Indian paneer. Paneer is made by coagulating milk with an acid or coagulant, and *W. coagulans* can be used as a natural coagulant in this process. The concentrated aspartic protease can be used instead of rennet for cheese preparation especially "cheddar". Cheese can be prepared by lyophilised extract has highest content of fat, total solids, crude protein, and ash which result in highest cheese yield as compared to pure chymosin and fungi rennet [18].

It's important to note that while *W. coagulans* can be used as a rennet substitute in cheese making, the specific techniques and processes may vary depending on the cheese recipe and desired outcome. Industrial cheese production often utilizes standardized rennet sources, but *W. coagulans* may find applications in small-scale or artisanal cheese production, especially in regions where it is traditionally used [19].

Overall, *W. coagulans* offers a natural alternative for coagulating milk in the dairy industry, particularly for those seeking vegetarian or vegan options in cheese making.

#### **8.1 Some key points highlighting the importance of** *W. Coagulans* **in the dairy industry**

Importance in the dairy industry due to its potential applications as a natural coagulant or rennet substitute in cheese and paneer production.


Ayurveda. The integration of traditional medicinal knowledge and practices into the dairy industry allows for a holistic approach to product development, incorporating both nutritional and therapeutic aspects.

While it has potential advantages, industrial-scale cheese production often utilizes standardized rennet sources. However, *W. coagulans* finds value in small-scale or artisanal cheese production and meets the demand for vegetarian and vegan options.

#### **9. The significance of** *W. coagulans* **in food industries**

#### **9.1 Nutritional profile**

Both macro and micronutrients are abundant in *W. coagulans*. The *W. coagulans* mineral constitution. In addition to being an excellent source of carbohydrates, it also contains a small amount of hydration, protein, fat, and fiber. Research studies have revealed that *W. coagulans* exhibits comparatively higher levels of essential minerals like magnesium (greater than *Alhagi maurorum*, *Berberis lyceum*, and *Tecomella undulate*), calcium (greater than *Dature alba*, *A. maurorum*, *Chenopodium album*, *B. lyceum*, *T. undulate*), potassium (greater than B. lyceum and T. undulate), and iron (greater than *D. alba*, *B. lyceum*, and *T. undulata*) [20]. The composition of roots, leaves, and fruit varies in terms of their nutrient content. Roots predominantly consist of carbohydrates (75.71%), followed by fiber (5.76%), lipids (5.5%), protein (2.95%), and ash (1.92%). In comparison, leaves contain carbohydrates (65.31%) as the major component, accompanied by fiber (11.76%), lipids (5%), protein (2.95%), and ash (3.26%). As for fruit, it consists of carbohydrates (60.14%) as the primary constituent, along with protein (4.65%), lipids (5%), and ash (4.21%). Hameed and Hussain [21]. The berries of *W. coagulans* include two esterases, free amino acids, milk coagulating enzymes, and essential oil. The primary amino acids found in the plant are proline, tyrosine, valine, hydroxyproline, glycine, cysteine, asparagine, glutamic, and aspartic acids. The major fatty acids found include arachidonic acid, stearic acid, palmitic acid, linoleic acid, and oleic acid [22]. Similarly, research found 20 constituents in the essential oil of the fruit of *W. coagulans* is primarily composed of sesquiterpenes (54%) and esters (21.50%), which are the dominant compounds. Additionally, fatty acids (5.5%) such as nonanoic acid, hexanoic acid, methyl ester of hexadecanoic acid, methyl ester of nondecanoic acid, methyl ester of 8,11-octadecadienoic acid, methyl ester of 9-octadecenoic acid, and ethyl ester of linoleic acid are present. Alkanes (9.11%) and aldehydes (0.32%) are present in smaller percentages Shahnaz *et al*. [23]. In addition, it was shown that a de-fatted meal made from *W. coagulans* seeds included free sugar (17.8%) in the form of D-galactose and D-arabinose (1:1), with trace amounts of maltose also present. Additionally discovered and reported for the hypocholesterolemic impact of corn oil combined with *W. coagulans* were higher concentrations of -sitosterol and linoleic acid. Maurya [11].

The fruit of the plant is known to contain certain constituents that may contribute to its potential health benefits. Here are some components found in *W. coagulans* fruit:

• *Withanolides*: *W. coagulans* is known to contain bioactive compounds called withanolides. These are steroidal lactones that are believed to have various pharmacological activities, including anti-inflammatory, antioxidant, and immunemodulating properties [24].

*Indian Cheese Revolution:* Withania coagulans *in Dairy Industry DOI: http://dx.doi.org/10.5772/intechopen.113815*


While detailed information on the specific nutrient composition of *W. coagulans* is limited, the following nutrients are commonly found in similar plants within the same family or genus:


The coagulating properties of *W. coagulans* berries on milk are widely recognized*.* Additionally, the milk coagulating potential of *W. coagulans* fruit extract was evaluated. *W. coagulans* fruit, an enzyme called aspartic protease was isolated using fractional ammonium-sulfate precipitation and cation-exchange chromatography. Casein was used to test the protease enzyme's proteolytic activity. Using skim milk, the ability of *W. coagulans* crude fruit extract to cause milk to coagulate was evaluated [17]. Therefore, it was discovered using mass spectrometry and inhibitory experiments that aspartic protease is the sole enzyme responsible for milk coagulation. Additionally, the activity of the enzyme was steadily decreased by the rising salt concentrations (NaCl, CaCl2). As a result, it was determined that this enzyme would be suitable to create reduced salt cheese [6].

Buffalo milk mozzarella cheese was made using the fruits of *W. coagulans*, which served as milk coagulants. Therefore, using an aqueous fraction of *W. coagulans* to make cheese might be an option. Buffalo milk cheese was made using an extract from the fruit of *W. coagulans*, and its storage capabilities were examined (5 months). Cheese produced using lyophilized berry extract showcased the highest


#### **Table 3.**

*Mineral composition of W. coagulans.*

concentrations of ash, fat, crude protein, and total solids. To create the cheese, *W. coagulans* alcoholic and aqueous fractions containing plant proteinase were utilized at varying concentrations (0.5, 1, and 1.5%) [28].

Additionally, cottage cheese made with an aqueous plant fraction had a significantly higher moisture content and pH, whereas cheese made with calf rennet and *W. coagulans* had the same levels of ash, fat, and crude protein. a white cheese of acceptable grade can be produced using a 0.5% alcoholic plant extract. Tofu made with calcium sulphate and *W. coagulans* extract were compared in terms of how well they produced coagulation of soy milk. The two types of tofu could not be distinguished based on sensory evaluation, while the tofu produced by *W. coagulans* had a lower yield and more moisture*.*

#### **10. Future prospects and potential of** *W. coagulans*

*W. coagulans*, also known as Indian rennet or vegetable rennet, is a medicinal plant that has been traditionally used in Ayurvedic medicine. General insights and potential avenues for exploration.


contribute to its health-promoting properties. Future studies may investigate the formulation of *W. coagulans* extracts or derivatives into various food products or supplements, evaluating their efficacy and safety profiles.


It's important to note that these prospects are speculative, and future research and developments will determine the actual potential and applications of *W. coagulans.*

### **Author details**

Mayur Ram1 \*, Bimal S. Desai<sup>2</sup> and Sumankumar S. Jha3

1 Department of Forest Products and Utilization, College of Forestry, Navsari Agricultural University, Navsari, Gujarat, India

2 Department of Basic Sciences and Humanities, College of Forestry, Navsari Agricultural University, Navsari, Gujarat, India

3 Department of Forest Biology and Tree Improvement, College of Forestry, Navsari Agricultural University, Navsari, Gujarat, India

\*Address all correspondence to: rammayur132@gmail.com

© 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.

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### *Edited by Adham M. Abdou*

Cheese is a palatable and dense model food with great nutritional value. Cheese lovers all over the world have access to an almost overwhelming variety of cheeses. Cheese has many health benefits that go beyond its nutritional and flavor properties. The health benefits are due to the presence of unique bioactive peptides and fatty acids in cheese. Furthermore, cheese is an excellent tool for making functional foods because it can serve as an excellent delivery vehicle for bioactive peptides, vitamins, minerals, probiotics, postbiotics, prebiotics, and other novel bioactive substances. This book offers opportunities for cheese manufacturers, cheese researchers, nutritionists, and even cheese lovers to learn more about the hidden health and nutritive benefits of cheese. The book reflects the trends and innovations in the development of cheese as a functional food.

> *Maria Rosário Bronze, Food Science and Nutrition Series Editor*

Published in London, UK © 2024 IntechOpen © Servet Turan / iStock

Recent Trends on Cheese as Functional Food with Great Nutritive and Health Benefits

IntechOpen Series

Food Science and Nutrition, Volume 4

Recent Trends on Cheese as

Functional Food with Great

Nutritive and Health Benefits

*Edited by Adham M. Abdou*