**Abstract**

Chemotherapeutic agents produce from numerous sources such as animals, plants and micro-organisms are derived from the natural products. Although the existing therapeutic pipeline lacks fungal-derived metabolites, but hundreds of secondary metabolites derived from fungi are known to be possible chemotherapies. Over the past three decades, several secondary metabolites such as flavonoids, alkaloids, phenolic and polyketides have been developed by *Saccharomyces cerevisiae* species with exciting activities that considered valued for the growth of new chemotherapeutic agents. Many secondary metabolites are protective compounds which prevent abiotic and biotic stresses, i.e. predation, infection, drought and ultraviolet. Though not taking part in a living cell's central metabolism, secondary metabolites play an important role in the function of an organism. Nevertheless, due to slow biomass build-up and inadequate synthesis by the natural host the yield of secondary metabolites is low by direct isolation. A detailed comprehension of biosynthetic pathways for development of secondary metabolites are necessary for *S. cerevisiae* biotransformation. These metabolites have higher inhibitory effect, specificity among cancer and normal cells, and the mechanism of non-apoptotic cell killing. This study shows the significance of bioactive compounds produced by *S. cerevisiae* species with their possible activity and value in chemotherapeutic drugs pipeline. The isolation and alteration of these natural secondary metabolites would promote the development of chemotherapeutic drugs.

**Keywords:** *Saccharomyces cerevisiae*, secondary metabolites, anticancer activity, synthetic biology, bioactive compounds

#### **1. Introduction**

Yeast is a single-celled eukaryotic microorganism that belongs to the kingdom of fungi. About 1500 yeast species have been correctly described since the discovery of the first yeast [1]. Yet 1% of all known fungal members are stated to be yeast species. *Saccharomyces cerevisiae*, budding yeas, also represents a typical industrial microorganism used in basic molecular biology research as a main model organism and was the first eukaryotic organism to have completely sequenced its genome.

A single-cell fungus, *Saccharomyces cerevisiae*, is also known as a Baker's yeast [2]. *Saccharomyces cerevisiae* is the eukaryotic microorganism most extensively studied, which allows us to understand the eukaryotic cell biology and subsequently the physiology of human. For several hundred years, in food processing and alcoholic drinks, *S. cerevisiae* have been used, and this organism is still used today in a variety of different pharmaceutical processes. It functions very well as it is non-pathogenic and is classified by GRAS organism (generally regarded as safe) due to the long history of use in the development of consumables such as ethanol and baker's yeast [3].

The sum of all organismic biochemical reactions can be described as metabolism. Metabolites are the products and intermediates of metabolism and are generally limited to small molecules. Natural products come from a variety of sources, including animal species, land-based plants, aquatic organisms, land invertebrates and vertebrates, microorganisms as secondary metabolite products [4]. The word "secondary" proposed by A. Kossel in 1891 means that while in any living cell primary metabolites are present, the secondary metabolites are only present at a byproduct and are of no significant importance to the life of the organism. Although the primary metabolism derives secondary metabolites, these secondary metabolites do not constitute the organism's fundamental molecular framework. Although the secondary metabolites do not participate in central metabolic processes of a living cell, they play an important role in the function of an organism. Many secondary bioactive compounds defend against biotic and abiotic and biotic stresses including predation, cancer, drought and ultraviolet radiations. Its absence does not reduce the life of an organism, which is a feature contrary to primary metabolite, instantly, but largely affected the organism' survival. Currently there are a range of analytical platforms for metabolomics research, among which are including mass spectrometry direct infusion (MS), gas chromatography liked to mass spectrometry (GC– MS), gas chromatography linking to mass spectrometry two-dimensional (GC, GC– MS), liquid chromatography coupled to mass spectrometry (LC–MS), capillary electrophoresis to mass spectrometry coupling (CE–MS), and proton nuclear magnetic resonance spectroscopy (1H NMR) [5].

These complex structural and chemical molecules serve as an excellent therapeutic class to cure many diseases. Around 80 percent of all drugs were derived from plant sources at the beginning of 1900. Alexander Fleming's discovery of penicillin from Penicillium notatum in 1928 marked a major change as a source of natural product from plants to micro-organisms. In medicine, agriculture, food industry and scientific research microorganism-derived compounds since then have been used [6]. But, due to the slow accumulation by biomass and inadequate synthesis by the host, the production of secondary metabolites by direct isolation is poor. Exhaustive awareness of biosynthetic pathways for the production of secondary metabolites are necessary for the development of *S. cerevisiae* biotransformation. Progress in synthetic biology has made it possible to develop a number of bioinformatics tools that can be utilized to create new regulatory elements and secondary metabolite synthesis pathways [7].

Because of its similar metabolism *Saccharomyces cerevisiae* was used as a biologically similar model for higher eukaryote organisms. The expression of heterogeneous pathways is less difficult relative to other kinds because of its eukaryotic nature. For example, prokaryotes do not conduct any post-translational modifications, and protein miss-folding and membrane translation can be an annoyance [8]. During preclinical or clinical testing, a well-characterized yeast metabolite, such as flavonoids, alkaloids, phenolic and polyketides, exhibit remarkable anti-tumor properties unexpectedly. Although its basic action mechanism is still being studied, evidence shows that its actions are operationally directed toward core regulatory pathways and dysregulated enzymes during cancer pathogenesis.

**111**

*Secondary Metabolites from* Saccharomyces cerevisiae *Species with Anticancer Potential*

Since ancient time, from the days of the Pharaohs in ancient Egypt, cancer continues to plague humanity. It derives its name from Hippocrates, father of medicine, who had been using the Greek-named "Karkinos" to talk about tumors, nevertheless these earliest view about this infection are different from modern concept. Cancer has been characterized as an irregular growth in cells caused by several changes in gene-expression leading to dysregulated equilibrium between death of cell and proliferation eventually developing into cell populations that can invade and metastasize tissues in distant sites and cause severe host death if left unsanctioned according to the World Health Organization (WHO). About 60 percent of all anticancer medications currently available in clinics are natural or derived from natural product modification substances. In the 1950s interest in the discovery and production of vinblastine, vinka alkaloids and vincristine, and later taxol from Pacific yew bark, *Taxus brevifolia*, in the pursuit of natural anticancer agents began earnestly. Then, our awareness of the metabolites of *S. cerevisiae* have also been further progressed in cancer therapy and have reported antitumor action against the overwhelming majority of cancers, like lymphoma, leukemia and solid tumors. Far from the suppression of tumor development, the delay of tumor progression, and an effect on tumor-cell metastatic and invading therapy, these metabolite combinations have almost all demonstrated strong therapeutic benefit at the preclinical level. Some of these allegedly promising metabolite compounds are

*DOI: http://dx.doi.org/10.5772/intechopen.95067*

discussed individually below [9].

**2. Production of secondary metabolites via** *S. cerevisiae*

metabolites are used to biosynthesize secondary metabolites [10].

particularly pharmaceutical products, from host (native) cells.

Apart from essential metabolites (carbons, proteins, amino acids, vitamins, acetones, ethanol, etc.), *S. cerevisiae* offers a wide range of secondary metabolites during active cell development, including toxins, antibiotics, fatty acids, alkaloids, alcohols, ketones etc. Secondary metabolites (SM's), which are not necessary for organism growth, are classified as diverse low molecular-weighted compounds. For a number of purposes, *S. cerevisiae* use secondary metabolites such as stress prevention, predation defense, competitiveness, communications, pathogenicity and exposure of other organisms. A small number of primary metabolism precursor

A number of bioactive compounds, including terpenoids, polyketides, alkaloids and non-ribosomal peptides can be produced in *Saccharomyces cerevisiae* (**Figure 1**). Such precursors are mainly short chain carboxylic acids (for example, acetyl-coA) or amino acids in *Saccharomyces cerevisiae* species which are connected to synthesized polyketides by backbone enzymes such as polyketide synthases (PKSs). Terpenoids production is catalyzed by terpene cyclase and production starting from dimethyl allyl diphosphate derived from (or isoprene). Non-ribosomal peptide synthetases (NRPSs) using modified and natural amino acids to synthesize non-ribosomal peptides. Diverse enzymes are used to catalyze the synthesis of alkaloids from amino acid. These secondary metabolites can naturally be synthesized as industrial products,

Many other reports have shown that the genes involved in biosynthetic secondary metabolites are known as biosynthetic gene clusters (BGCs). This have been shown that genes that encode significant biosynthesis (for example; polyketides synthase) are supplementary enzymes and precursors in the pathways of biosynthesis. Consequently, all significant genes are discovered in the BGCs that participate in the synthesis of bioactive compounds [11, 12]. The production and complementary regulation of catalytic properties can produce incredibly useful secondary metabolites for these biochemical transformations BGC enzymes [13].

*Secondary Metabolites from* Saccharomyces cerevisiae *Species with Anticancer Potential DOI: http://dx.doi.org/10.5772/intechopen.95067*

Since ancient time, from the days of the Pharaohs in ancient Egypt, cancer continues to plague humanity. It derives its name from Hippocrates, father of medicine, who had been using the Greek-named "Karkinos" to talk about tumors, nevertheless these earliest view about this infection are different from modern concept. Cancer has been characterized as an irregular growth in cells caused by several changes in gene-expression leading to dysregulated equilibrium between death of cell and proliferation eventually developing into cell populations that can invade and metastasize tissues in distant sites and cause severe host death if left unsanctioned according to the World Health Organization (WHO). About 60 percent of all anticancer medications currently available in clinics are natural or derived from natural product modification substances. In the 1950s interest in the discovery and production of vinblastine, vinka alkaloids and vincristine, and later taxol from Pacific yew bark, *Taxus brevifolia*, in the pursuit of natural anticancer agents began earnestly. Then, our awareness of the metabolites of *S. cerevisiae* have also been further progressed in cancer therapy and have reported antitumor action against the overwhelming majority of cancers, like lymphoma, leukemia and solid tumors. Far from the suppression of tumor development, the delay of tumor progression, and an effect on tumor-cell metastatic and invading therapy, these metabolite combinations have almost all demonstrated strong therapeutic benefit at the preclinical level. Some of these allegedly promising metabolite compounds are discussed individually below [9].

### **2. Production of secondary metabolites via** *S. cerevisiae*

Apart from essential metabolites (carbons, proteins, amino acids, vitamins, acetones, ethanol, etc.), *S. cerevisiae* offers a wide range of secondary metabolites during active cell development, including toxins, antibiotics, fatty acids, alkaloids, alcohols, ketones etc. Secondary metabolites (SM's), which are not necessary for organism growth, are classified as diverse low molecular-weighted compounds. For a number of purposes, *S. cerevisiae* use secondary metabolites such as stress prevention, predation defense, competitiveness, communications, pathogenicity and exposure of other organisms. A small number of primary metabolism precursor metabolites are used to biosynthesize secondary metabolites [10].

A number of bioactive compounds, including terpenoids, polyketides, alkaloids and non-ribosomal peptides can be produced in *Saccharomyces cerevisiae* (**Figure 1**). Such precursors are mainly short chain carboxylic acids (for example, acetyl-coA) or amino acids in *Saccharomyces cerevisiae* species which are connected to synthesized polyketides by backbone enzymes such as polyketide synthases (PKSs). Terpenoids production is catalyzed by terpene cyclase and production starting from dimethyl allyl diphosphate derived from (or isoprene). Non-ribosomal peptide synthetases (NRPSs) using modified and natural amino acids to synthesize non-ribosomal peptides. Diverse enzymes are used to catalyze the synthesis of alkaloids from amino acid. These secondary metabolites can naturally be synthesized as industrial products, particularly pharmaceutical products, from host (native) cells.

Many other reports have shown that the genes involved in biosynthetic secondary metabolites are known as biosynthetic gene clusters (BGCs). This have been shown that genes that encode significant biosynthesis (for example; polyketides synthase) are supplementary enzymes and precursors in the pathways of biosynthesis. Consequently, all significant genes are discovered in the BGCs that participate in the synthesis of bioactive compounds [11, 12]. The production and complementary regulation of catalytic properties can produce incredibly useful secondary metabolites for these biochemical transformations BGC enzymes [13].

*Saccharomyces*

A single-cell fungus, *Saccharomyces cerevisiae*, is also known as a Baker's yeast [2]. *Saccharomyces cerevisiae* is the eukaryotic microorganism most extensively studied, which allows us to understand the eukaryotic cell biology and subsequently the physiology of human. For several hundred years, in food processing and alcoholic drinks, *S. cerevisiae* have been used, and this organism is still used today in a variety of different pharmaceutical processes. It functions very well as it is non-pathogenic and is classified by GRAS organism (generally regarded as safe) due to the long history of use in the development of consumables such as ethanol and baker's yeast [3]. The sum of all organismic biochemical reactions can be described as metabolism. Metabolites are the products and intermediates of metabolism and are generally limited to small molecules. Natural products come from a variety of sources, including animal species, land-based plants, aquatic organisms, land invertebrates and vertebrates, microorganisms as secondary metabolite products [4]. The word "secondary" proposed by A. Kossel in 1891 means that while in any living cell primary metabolites are present, the secondary metabolites are only present at a byproduct and are of no significant importance to the life of the organism. Although the primary metabolism derives secondary metabolites, these secondary metabolites do not constitute the organism's fundamental molecular framework. Although the secondary metabolites do not participate in central metabolic processes of a living cell, they play an important role in the function of an organism. Many secondary bioactive compounds defend against biotic and abiotic and biotic stresses including predation, cancer, drought and ultraviolet radiations. Its absence does not reduce the life of an organism, which is a feature contrary to primary metabolite, instantly, but largely affected the organism' survival. Currently there are a range of analytical platforms for metabolomics research, among which are including mass spectrometry direct infusion (MS), gas chromatography liked to mass spectrometry (GC– MS), gas chromatography linking to mass spectrometry two-dimensional (GC, GC– MS), liquid chromatography coupled to mass spectrometry (LC–MS), capillary electrophoresis to mass spectrometry coupling (CE–MS), and proton

nuclear magnetic resonance spectroscopy (1H NMR) [5].

secondary metabolite synthesis pathways [7].

pathways and dysregulated enzymes during cancer pathogenesis.

These complex structural and chemical molecules serve as an excellent therapeutic class to cure many diseases. Around 80 percent of all drugs were derived from plant sources at the beginning of 1900. Alexander Fleming's discovery of penicillin from Penicillium notatum in 1928 marked a major change as a source of natural product from plants to micro-organisms. In medicine, agriculture, food industry and scientific research microorganism-derived compounds since then have been used [6]. But, due to the slow accumulation by biomass and inadequate synthesis by the host, the production of secondary metabolites by direct isolation is poor. Exhaustive awareness of biosynthetic pathways for the production of secondary metabolites are necessary for the development of *S. cerevisiae* biotransformation. Progress in synthetic biology has made it possible to develop a number of bioinformatics tools that can be utilized to create new regulatory elements and

Because of its similar metabolism *Saccharomyces cerevisiae* was used as a biologically similar model for higher eukaryote organisms. The expression of heterogeneous pathways is less difficult relative to other kinds because of its eukaryotic nature. For example, prokaryotes do not conduct any post-translational modifications, and protein miss-folding and membrane translation can be an annoyance [8]. During preclinical or clinical testing, a well-characterized yeast metabolite, such as flavonoids, alkaloids, phenolic and polyketides, exhibit remarkable anti-tumor properties unexpectedly. Although its basic action mechanism is still being studied, evidence shows that its actions are operationally directed toward core regulatory

**110**

#### **Figure 1.**

*A short description of the formation pathways of secondary metabolites [11, 12]. Four sub-groups of microorganism and plant-generative metabolites involve terpenoids, polyketides, non-ribosomal peptides and alkaloids. Biosynthetic genes (BGC's) are structurally classified into the genes that encrypt major biosynthetic enzymes, supplementary enzymes, precursors in the process of biosynthesizing, resistance factors or transporters in secondary metabolite formation.*

The methods and technology used to improve pharmaceutical production of secondary metabolites in the *S. cerevisiae* have advanced rapidly in recent years.

We discuss below the progress in developing biosynthetic pathways of *Saccharomyces cerevisiae*' secondary metabolites with anticancer potential: alkaloids, phenolic, terpenoids, polyketides, non-ribosomal peptides, and vitamin C.

## **3. Alkaloids**

Several studies have used *S. cerevisiae* in recent years as a host to engineer the biosynthetic process of alkaloids. Alkaloids are complex nitrogen molecules that are extremely bioactive. There are reportedly approximately 50 alkaloid medications, including vincristine (cancer drug), codeine (analgesic drug) and noscapine (antitussive drug). They are unique for the cell cycle and the process, because they block metaphase (M phase) in mitosis. They block tubulin's ability to form microtubules via polymerization. Dysfunctional spindle structures, avoid chromosome separation and cell proliferation. Strictosidine, which had been synthesized by novo, was the earliest study of alkaloid plant origin in *Saccharomyces cerevisiae* strains [14]. Strictosidine is a significant intermediate product in the terpenoids Indole alkaloids (TIAs) biosynthesis that include vincristine (antitumor) and ajmalicine (antihypertensive) [15].

The metabolic course of *Saccharomyces cerevisiae* for protoberberine alkaloid (S) canadine from racnorlaudanosolin has been optimized by Galanie et al. [16]. The secondary metabolite of Berberine has numerous pharmacological effects, such as antidiabetics, antibacterial, anti-ulcerones and anti-inflammatory effects. In vivo and vitro experiments with berberine, the results of arresting cell cycle during G1 or G2/M and apoptosis of tumor cell were shown to be anti-cancer activity. Berberine also was found to induce autophagy and stress in the endoplasmic reticulum, resulting in the invasion and inhibition of the tumor cells. Berberine has been proven to lower angiogenesis by decreasing expression of VEGF in addition to its apoptotic effects. The cancer cell migration was also decreased. In the respective

**113**

*Secondary Metabolites from* Saccharomyces cerevisiae *Species with Anticancer Potential*

complexes, Berberine attaches to DNA or RNA. Berberine also stimulates P53, which contributes to cell cycle arrest and apoptosis. Berberine has been shown to induce apoptosis as well via DNA interactions and pathways that are dependent on

Phenolic are a large category of secondary plant metabolites that have at least one hydroxylated ring. Flavonoids and stilbenes are two groups of metabolites highly appreciated as a nutrient and therapeutic agent among the phenolic developed by phenylpropanoid pathways in the plant [18]. As a host cell, *Saccharomyces cerevisiae* is well equipped to promote phenolic compound biosynthesis. The following phases include cancer development: initiation, development, proliferation, invasion and metastasis. Initiation links by free radicals, inflammatory mediators, radiation, smoke of cigarette that damage the DNA-product, which can cause genetic mutation and replication of cells which are mutated results to cause carcino-

It has been noted that phenolic compounds i.e. ellagic acid and delphinidin show significant protective effects for example, apoptotic activity in colon cancer cells. In prostate, liver and leukemia cancer cells, delphinidin also showed significant activity. Cell apoptosis can also be caused by phenolic compounds. The mechanisms

of invasion and migration of human cancer are also updated (**Figure 2**) [20].

The *Saccharomyces cerevisiae* strains have been developed by researchers, which synthesize various flavonoids, from the main phenolic intermediates to flavones, flavanones, flavonols and isoflavones. Phenylalanine is transformed into p-coumaric acid, a common phenylpropanoid precursor, in two sequence reactions in the initial steps of the phenylpropanoid pathway. *S. cerevisiae* is an ideal flavonoid host strain. One of the first studies on *S. cerevisiae* for flavonoid production was carried out according to [21]. The first flavonoid analysis was published on the synthesis of naringenin, major intermediate of flavonoid, from glucose in engineered *Saccharomyces cerevisiae* [22]. In *Saccharomyces cerevisiae* p-coumaric acid was used as a precursor for other flavonoids including apigenin, chrysin and luteolin. Anti-inflammatory, cardio-protective and anticancer effects were

*Potential of secondary metabolite protective mechanisms for cancer management [17].*

observed for quercetin, kaempferol, and fisetin [23].

*DOI: http://dx.doi.org/10.5772/intechopen.95067*

mitochondrial [17].

**4. Phenolic**

genesis [19].

**5. Flavonoids**

**Figure 2.**

*Secondary Metabolites from* Saccharomyces cerevisiae *Species with Anticancer Potential DOI: http://dx.doi.org/10.5772/intechopen.95067*

complexes, Berberine attaches to DNA or RNA. Berberine also stimulates P53, which contributes to cell cycle arrest and apoptosis. Berberine has been shown to induce apoptosis as well via DNA interactions and pathways that are dependent on mitochondrial [17].
