Fermentation for Peptides

As a leading CDMO, BOC Sciences provides one-stop fermentation services to produce custom peptide for various industries such as pharmaceutical, food, nutraceutical and cosmetic. Our manufacturing systems comply with good manufacturing practice (GMP), which are well suited to meet the needs of our customers around the world.

Learn more about Fermentation Products.

What are Bioactive peptides?

Bioactive peptides are a very useful family of functional molecules with several health benefits. These chemicals are released in a variety of fermented foods made with different starting microorganisms. Bioactive peptides can be used as strong alternatives to synthetic molecules as components in nutraceuticals and functional food items due to their potential high bioactivity and low negative effects. Peptide fermentation is a biotechnological technique that produces bioactive peptides through microbial fermentation processes. Peptides offer a wide range of uses, including medications, cosmetics, dietary supplements, and agricultural goods. The proteolytic specificity of lactic acid bacteria, Bacillus spp., yeasts, and mold, in addition to their capacity to manufacture bioactive peptides, is significant in the creation of particular bioactive peptides in traditional fermented foods.

Fermentation Production of Peptides

In microbial fermentation method, the substrate protein is hydrolyzed by the protease produced during the growth of the bacterial strain. Then the fermentation product is separated and purified, and finally the bioactive peptide is obtained. The production process technology of microbial fermentation is mainly to convert biological macromolecule protein into small molecule peptide according to contemporary microbial fermentation technology. Peptides with different amino acid arrays and molecular weights can be produced according to the metabolic and alcoholization standards by manipulating microorganisms.

Advantages of Fermentation for Peptides

Fermentation is a one-step process to prepare polypeptides. The preparation of polypeptides by microbial fermentation has the advantages of low cost, high safety, stable quality and remarkable function. The peptides produced by microorganisms have been proved to have numerous beneficial health effects, especially in food industry.

Application of Fermentation for Peptides

In the whole process of alcoholization, the resulting dispersed amino acids are digested, absorbed and used by microorganisms again, which is not easy to cause feedback inhibition to the metabolism of microorganisms. According to microbial metabolism, amino acids and small peptides were grafted and rearranged, and some peptide functional groups were decorated and assets were reorganized. For example, soybean peptides produced by microbial fermentation with soya bean as raw materials changed the original amino acid sequence of soybean protein, decorated the hydrophobic amino acid tail end of the peptide, made the soybean peptides not bitter taste, higher activity, and granted some biological activities of soybean peptides.

In addition, antimicrobial peptides produced by fermentation have also made progress in recent years. Antimicrobial peptides are a class of peptides that resist the defense response of microorganisms. It has the characteristics of small molecular weight, good water solubility, strong heat resistance, no immunogenicity, and wide antibacterial spectrum. Antimicrobial peptides are peptides encoded by specific genes in animal and plant cells and induced by external conditions to kill microorganisms such as bacteria, fungi and viruses. The antimicrobial peptides produced by microorganisms isolated from fermented foods are generally harmless to humans. Therefore, it is safer and more effective than many preservatives when added to medicine and food as additives. Moreover, because of its low dosage, it is not easy to make organisms develop resistance.

Fermented peptide from dairy products

Milk is abundant in nutritionally valuable proteins (casein and whey proteins), which account for up to 3-6% of total milk composition. The sequence of milk proteins encodes many bioactive fragments, which are released as peptides when milk is hydrolyzed by starter strains during the fermentation process. Milk is an ideal substrate for LAB development, and LAB proteolytic systems hydrolyze milk proteins to produce a variety of bioactive peptides. Lactobacillus spp., Lactococcus spp., Lactiplantibacillus spp., Lacticaseibacillus spp., Leuconostoc spp., Streptococcus spp., and Enterococcus spp. proteolytic systems have resulted in the release of milk-derived peptides with antioxidant, antihypertensive, antidiabetic, and immunomodulatory properties. After being fermented for 24 hours at room temperature with kefir probiotic bacteria, the milk sample was analyzed and found to contain around 35 peptides, including the β-CN-derived peptide YQEPVLGPVRGPFPIIV, which has been shown to have antihypertensive properties in vivo. After being matured for more than 180 days, skin bag Tulum cheese from Turkey had 203 different peptides. After more investigation, bioactive peptides with antihypertensive, antibacterial, antioxidant, immunomodulatory, anti-inflammatory, and anti-carcinogenic qualities were found in the Tulum cheese made the old-fashioned way. This supports the idea that for milk proteins to produce bioactive peptides, fermentation and ripening are essential.

Fermented peptide from meat and fish products

Excellent sources of high-nutrient-value proteins in our diet include meat and fish. The breakdown of proteins, lipids, and carbohydrates into flavor-enhancing end products by fermenting starter strains and endogenous muscle enzymes is responsible for the meat products' enhanced sensory appeal.  The two most common proteolytic strains linked to the manufacturing of European fermented meat products are staphylococci and LAB. In addition to these bacterial strains, several Asian nations also utilize the koji mold, or Aspergillus oryzae, to produce fermented meat sauce.

During the ripening phase of dry-cured hams, microbial enzymes hydrolyze meat proteins, producing peptides with useful characteristics. The 8–10 month ripened ham has been discovered as a possible source of antioxidant peptides due to the presence of antioxidant peptides such LPGGGHGDL and LPGGGT, among the 33 peptides found in Chinese dry-cured Jinhua ham. Peptide fractions of European hams, such as Belgian dry-cured hams, Italian Parma, and Spanish Teruel hams, showed antioxidant and ACE-inhibition properties. Possible bioactive peptides with antioxidant and ACE-inhibitory properties were found in camel sausage that was fermented by Lactobacillus pentosus, Lactobacillus sakei, and Staphylococcus xylosus.

Protein hydrolysates of scallop (Argopecten irradians) mantle fermented with Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloliquefaciens revealed the discovery of novel antioxidant peptides (ALLEEWEK and KLADMLNPER).

Generation of bioactive peptides during the production of diverse fermented food products. (Chourasia R., et al., 2023)

Food derived peptides as potential therapeutics against β-CoVs

Since the SARS-CoV outbreak, there has been a great deal of research done on the immunostimulatory and viral suppressive properties of both natural and artificial peptides. High antiviral activity peptides obtained from food can be utilized to create medicines that effectively combat β-CoVs. In silico studies have shown that antiviral peptides generated during microbial fermentation and enzymatic degradation of dietary proteins potently block β-CoV attachment and reproduction. Notwithstanding these results, more thorough research and development work on peptide-based candidates are required to provide potent medicines that are especially effective against β-CoV infections.

Peptides generated from food have been shown to interact with NSPs and β-CoV structural proteins, potentially inhibiting viral infection and growth. Food fermentation releases bioactive peptides, which have a variety of uses such as antibacterial, antioxidant, antihypertensive, and anticancer effects. These peptides can be investigated further to create nutraceuticals and medications. Peptides from fermented food have shown strong antiviral efficacy against viruses in the past. Based on soy cheese made with Lactobacillus delbrueckii WS4, the peptide KFVPKQPNMIL showed strong binding affinity to important residues of S1 RBD and 3CLpro of SARS-CoV-2, SARS-CoV, MERS-CoV, and HCoV-HKU1. This suggests that the peptide may be able to inhibit the attachment and replication of β-CoVs.

These food-derived peptides have the ability to interact with many viral proteins, making them useful as lead molecules for the development of effective anti-β-CoV therapies. The soybean fermentation process produced the peptide ALPEEVIQHTFNLKSQ, which exhibited a strong affinity for both S1 RBD and TLR4/MD2 complex. This was determined by conducting molecular docking analyses of peptides derived from fermented soybeans against SARS-CoV-2 RBD and human TLR4/Myeloid Differentiation factor 2 (MD2) complex.

Molecular docking experiments employing peptides derived from in silico gastrointestinal (GI) digestion of wheat, barley, and oat proteins revealed high-affinity interaction with SARS-CoV-2 S1 RBD. In another in silico analysis, the peptide VPW produced from edible mealworms shown a higher binding affinity to SARS-CoV-2 RBD than certain natural products. During molecular docking investigations, in silico GI digestion of storage proteins from quinoa, sesame, rape, sunflower, and pumpkin seeds resulted in the release of numerous peptides with high GI absorption and binding affinities to several structural proteins and NSPs of SARS-CoV-2. Peptides derived from in silico GI digestion of marine fish proteins showed a strong affinity for critical catalytic residues of SARS-CoV-2 3CLpro. The tuna skeletal myosin-derived peptide EEAGGATAAQIEM has strong water solubility, negligible toxicity, and a high binding affinity for key 3CLpro residues, including the HIS41-CYS145 catalytic dyad. Food-derived peptides can suppress viral entrance and replication, making them potential treatments and prophylactics for β-CoV illnesses.

Potential anti-β-CoV activities of bioactive peptides from microbial fermentation. (Chourasia R., et al., 2022)

What Can We Do?

BOC Sciences has the ability to convert a variety of traditional strains into engineered strains and provide one-stop fermentation CDMO services from laboratory scale to industrial scale fermentation services. Our fermentation product list includes proteins, industrial enzymes, and other small molecule compounds, such as peptides.

References

1. Chai KF. et al. Bioactive peptides from food fermentation: A comprehensive review of their sources, bioactivities, applications, and future development. Compr Rev Food Sci Food Saf. 2020, 1-61.
2. Lazzaro BP. et al. Antimicrobial peptides: Application informed by evolution. Science. 2020, 368(6490).
3. Chourasia R., et al., Bioactive peptides in fermented foods and their application: A critical review, Systems Microbiology and Biomanufacturing, 2023, 3(1): 88-109.
4. Chourasia R., et al., Peptide candidates for the development of therapeutics and vaccines against β-coronavirus infection, Bioengineered, 2022, 13(4): 9435-9454.
5. Chai K F., et al., Bioactive peptides from food fermentation: A comprehensive review of their sources, bioactivities, applications, and future development, Comprehensive Reviews in Food Science and Food Safety, 2020, 19(6): 3825-3885.