Unlocking the Power of Fibrinolytic Enzymes in Traditional Indian Fermented Foods: A Gateway to Natural Antithrombotic Agents

Applying food enzymes in the kitchen

Ensuring food safety and at the same time meeting such demands for retention of nutrition and quality attributes have resulted in increased interest in alternative preservation techniques for inactivating microorganisms and enzymes in foods. This increasing demand has opened new dimensions for the use of natural preservatives derived from plants, animals, or microflora. Extensive research has investigated the potential application of natural antimicrobial agents in food preservation. Especially the significance and use of microbes as producers of antimicrobial metabolites has increased significantly during the last decades. Reported studies have demonstrated that microbial metabolites from microorganisms exhibited a great numbers of diverse and versatile biological effects about antimicrobial activities. These microorganisms produce many compounds that are active against other microorganisms, which can be harnessed to inhibit the growth of potential spoilage or pathogenic microorganisms. These include fermentation end products (metabolites) such as organic acids, hydrogen peroxide, and diacetyl, biofilm, exopolysaccharides in addition to bacteriocins and other antagonistic compounds such as reuterin. Up to now, antimicrobial metabolites from lactic acid bacteria (such as nisin) have been mostly used in food preservation. In addition to lactic acid bacteria, some yeast, mold, and another bacteria species as well as some pathogenic bacteria can produce antimicrobial metabolites. Antimicrobial metabolites present in foods can extend the shelf life of unprocessed or processed foods by reducing the microbial growth rate or viability. This offers a new knowledge-based approach to the exploitation of bacteria for food production, from metabolic engineering of microorganisms to produce antimicrobials or nutritionals, to the molecular mining of activities as yet unknown but which could benefit food production. In addition, the availability of the genomes of many food pathogenic and spoilage bacteria may open up new possibilities for the design of novel antimicrobials which target essential functions of these problematic bacteria. In this chapter, antimicrobial metabolites from microorganism in food safety as a biocontrol agent reviewed.

Fibrinolytic enzymes in Asian traditional fermented foods

Protective, technological, and functional properties of select autochthonous lactic acid bacteria from goat dairy products

Boosting immunity is the most important preventive measure to fight against different externally acquired infection like SARS-COV2 outbreak presently prevailing all over the world. Food plays a major role in enhancing immunity of the body. Fermented foods especially fermented milk foods are a great source of bioactive peptides that help in boosting immunity. India has a plethora of traditional fermented milk foods such as dahi, lassi, srhikhand, chhurpi etc. in its kitty which are having abundant biofunctional activities. They exhibit various health benefits especially immunomodulation, antimicrobial, antithrombotic and antioxidative properties which are mostly required to combat the detrimental effects of the pathogens. They are also a rich source of several vitamins and micronutrients required for boosting immunity. The present article focuses on the biofunctional properties of traditional Indian fermented milk foods which have the capability of enhancing immunity of the body. It also explores the way to fight against the SARS-COV2 virus by marinating proper immunity with simple available resources found in traditional Indian fermented milk foods.

The concept of "ethno-microbiology" is to understand the indigenous knowledge of the Indian people for production of culturally and organoleptically acceptable fermented foods by natural fermentation. About 1000 types of common, uncommon, rare, exotic and artisan fermented foods and beverages are prepared and consumed in different geographical regions by multi-ethnic communities in India. Indian fermented foods are mostly acidic and some are alkaline, along with various types of alcoholic beverages. A colossal diversity of microorganisms comprising bacteria mostly belongs to phylum Firmicutes, filamentous moulds and enzyme- and alcohol-producing yeasts under phyla Ascomycota and Mucoromycota, and few bacteriophages and archaea have been reported from Indian fermented foods. Some microorganisms associated with fermented foods have functionalities and health promoting benefits. "Ethno-microbiology" of ethnic Indian people has exhibited the proper utilisation of substrates either singly or in combination such as fermented cereal-legume mixture (idli, dosa and dhokla) in South and West India, sticky fermented soybean food (kinema and related foods), fermented perishable leafy vegetable (gundruk and related foods), fermented bamboo shoots (soibum and related foods) and fermented fish (ngari and others) in North East India, and fermented meat and sausage-like products in the Indian Himalayas, fermented coconut beverage (toddy) in coastal regions, and various types of naturally fermented milk products (dahi and related products) in different regions of India. This review has also highlighted the "ethno-microbiology" knowledge of the people involving the consortia of essential microorganisms in traditionally prepared amylolytic starters for production of cereal-based alcoholic beverages. The novelty of this review is the interpretation of ethno-microbiological knowledge innovated by ethnic Indian people on the use of beneficial microorganisms for food fermentation to obtain the desired fermented food products for consumption.

Fermentation Technology and Food Enzymes

Cardiovascular disease is among the largest contributors of premature mortality in the world caused by inflammation of blood vessels. The abnormalities provoke thrombus formation or thrombosis blocking blood vessels leading to strokes, heart attacks and coronary artery diseases. Increasing percentage of cardiovascular cases and deaths due to thrombosis has attracted researchers to look for newer thrombolysis agents. Commonly used drugs to treat thrombosis has been limited due to various side effects. Therefore, the search for sources of safer and cheaper fibrinolytic enzymes for handling thrombolysis continues. This study aimed to evaluate potentials of fibrinolytic protease of bacteria isolated from fermented seafood (sea cucumber) products as antithrombotic agents. Information was initially gathered from scientific publications identified using web-based tools including PubMed (National Center for Biotechnology Information), Science Direct (Scopus) and Web of Science (Thomson Reuters) using combinations of search terms including “fibrinolytic enzyme protease”, “endopeptidase”, “fermented food”, “sea cucumber”, “thrombolysis therapy,” “thrombolytic agent,” “fibrinolytic bacteria,” “fibrinolysis,” “protease producing bacteria,” “fibrin degradation,” “holothurians,” etc. We also searched for these terms in national and international organization technical reports and databases. This literature review reveals the prospects of fibrinolytic protease enzymes from bacteria from fermented seafood, particularly sea cucumber as novel antithrombotic agents.

<em><span>Data collected by WHO in 2019 shows an estimated 17.9 million (equivalent to 32% of global death). The total number is due to cardiovascular disease, 85% were due to heart attacks and strokes. One of the basic pathophysiologies of stroke is atero-thromboembolic (thrombosis). The accumulation of clots formed by fibrin in blood vessels can inhibit several processes in the body system, such as blood flow, oxygen transportation, and transportation of nutritional and non-nutritional compounds needed. Fibrinolytic enzymes from food microbes have attracted attention for further research as thrombolytic agents. The Bacillus genus from fermented foods can produce strong fibrinolytic enzymes, such as Bacillus natto from natto (Japan), Bacillus subtilis KCK-7 from Chungkook-jang (Korea), Bacillus weihenstephanensis from shrimp paste (Vietnam), Bacillus amyloliquefaciens MH18B1 from Hawaijar (India), Rhizopus chinensis 12 from Chinese wine (China), and Bacillus pumillus and Bacillus subtilis K2 from fermented soybean food products such as tempe gembus, red oncom, and moromi (Indonesia). The high protein content in soybeans can be used as a medium for the growth of proteolytic and fibrinolytic microorganisms. Studies in Asia regarding fibrinolytic enzymes from fermented foods are still limited, even though the Asian region comprises many countries rich in fermented foods. This review is relevant to healthy food ingredients from local fermented foods. Recently, microbial fibrinolytic enzymes from fermented foods have received tremendous interest in the medical field. Many local fermented foods are a potential source of fibrin-degrading enzymes, which can be further developed to treat diseases like stroke and heart attack. This article aims to review microbial fibrinolytic enzymes from various fermented foods and their impact on health, especially thrombosis. This manuscript review uses the narrative review method. From the overall discussion, it can be concluded that fermented foods have properties as fibrinolytic agents.</span></em>

Ethnic Indian people have been domesticating beneficial microorganisms (bacteria, yeasts, and moulds) by their wisdom of ethno-microbiological knowledge for production of flavoured and socio-culturally preferred fermented foods and alcoholic beverages for more than 8000 years. The purpose of this review is to collate the available literatures of diversity of Saccharomyces and non-Saccharomyces species associated with Indian fermented foods and alcoholic beverages. A colossal diversity of enzyme- and alcohol-producing yeasts under the phylum Ascomycota has been reported from Indian fermented foods and alcoholic beverages. The distributions of yeast species show 13.5% of Saccharomyces cerevisiae and 86.5% of some non-Saccharomyces spp. in Indian fermented foods and alcoholic beverages, based on the reported literatures available till date. There is a research gap on prospect of yeasts research in India. Hence, we suggest that validation of traditional knowledge of domestication of functional yeasts needs to be studied to develop the functional genomics platforms for Saccharomyces and non-Saccharomyces spp. in Indian fermented foods and alcoholic beverages.

Biosynthesis of Enzymes in Fermented Foods by Lactic Acid Bacteria and Possible Use as Bio-Preservative in Fermented Milk

This article provides an overview of the characteristics, applications, and potential of Aspergillus niger in dietary enzyme production. A. niger is a filamentous fungus that grows naturally in a wide variety of temperatures and pH levels. It is frequently utilized in the synthesis of citric acid and other organic acids. A. niger is also a significant organism in traditional fermented foods, and it has been utilized to make food enzymes that have been designated as Generally Recognized As Safe (GRAS) by the United States Food and Drug Administration. The constraints of A. niger genetic manipulation are discussed, as well as the need to eradicate mycotoxins in industrial strains. The article finishes with an overview of current advances in genetic manipulation and enzyme overproduction tactics, which may assist to increase the efficiency and safety of A. niger as a potential enzyme host in the food industry

Fermented foods as biotechnological resources

This review describes the diversity of Indian fermented food and its significance as a potential source of lactic acid bacteria (LAB). Fermented foods consumed in India are categorized based upon their base material. Fermented foods such as dahi, gundruk, sinki, iniziangsang, iromba, fermented rai, kanjika and handua were reported to have significant medicinal properties. Some fermented products such as koozh, dahi and kanjika are consumed unknowingly as, probiotic drinks, by local people. There are very few reports regarding isolation of LAB from Indian fermented foods available in the past; however, due to growing consciousness about potential health benefits of LAB, we now have scores of reports in this field. There is an abundant opportunity available for food microbiologists to explore the Indian fermented foods for the isolation of new LAB strains for their potential role in probiotic research.

Lactic acid bacteria (LAB) were isolated from naturally fermented foods of India, viz., sidra, a dried fish product; kinema, a naturally fermented sticky soybean food; and dahi, a naturally fermented milk product. Five strains of LAB, based on 16S rRNA gene sequence, were identified: Lactococcus lactis FS2 (from sidra), Lc. lactis C2D (dahi), Lc. lactis SP2C4 (kinema), Lactiplantibacillus plantarum DHCU70 (=Lactobacillus plantarum) (from dahi), and Lactiplantibacillus plantarum KP1 (kinema). The PICRUSt2 software, a bioinformatic tool, was applied to infer the raw sequences obtained from LAB strains mapped against KEGG database for predictive functionality. Functional features of LAB strains showed genes associated with metabolism (36.47%), environmental information processing (31.42%), genetic information processing (9.83%), and the unclassified (22.28%). KEGG database also showed abundant genes related to predictive membrane transport (29.25%) and carbohydrate metabolism (11.91%). This study may help in understanding the health-promoting benefits of the culturable LAB strains in fermented foods.