Isolation, production and application of fibrinolytic enzyme from fermented food sources

2021 ◽  
Author(s):  
Nitika Singh ◽  
Shailendra Singh Shera
Keyword(s):  
Cardiovascular Diseases ◽  
Blood Clot ◽  
Fermented Food ◽  
Fibrinolytic Enzyme ◽  
Food Sources ◽  
Bacillus Species ◽  
Cow Dung ◽  
Fermented Foods ◽  
Fibrinolytic Enzymes ◽  
Wide Range

The accumulation of the fibrin in bacterial fibrinolytic enzymes finds applications to treat and prevent cardiovascular diseases which fail in hemostasis that leads to the formation of undesirable blood clots in the blood vessels leading to condition called thrombosis. The fibrinolytic enzymes from food grade organisms are useful for thrombolytic therapy. Conventional thrombolytic agents such as streptokinase, nattokinase etc.  Nattokinase is one such fibrinolytic enzyme with a wide range of applications in Pharmaceutical industry, health care and medicine etc. Hence, potent blood-clot dissolving protein used for the treatment of cardiovascular diseases is produced by the bacterium Bacillus subtilis during the fermentation of soybeans to produce Natto. The health benefits of some fermented foods are synthesis of nutrients, prevention of cancer, diabetes due to presence of functional microorganisms, which possess probiotics properties, antimicrobial, antioxidant, etc. The first report of fibrinolytic enzyme production of cow dung used as a cheap substrate from Bacillus species in SSF has been given earlier. This review describes different isolation methods which enable the screening and selection of promising organisms for industrial production. The purification and properties of these fibrinolytic proteases is discussed, and the use of fibrinolytic enzyme. In order to obtain Bacillus species producing fibrinolytic enzymes, the fermented food sample such as sprouted grain and processed grain etc were used. The heat tolerant isolates initially were selected for catalase test. Fibrinolytic activity of the selected isolates was determined by using Fibrin plate assay.  From the above work, it can be concluded that the fibrinolytic enzyme produced by Bacillus from fermented food samples had the ability to degrade the fibrin and hence can be used for functional food formulation.

scholarly journals PRODUCTION AND PARTIAL CHARACTERIZATION OF FIBRINOLYTIC ENZYME FROM A SOIL ISOLATE ASPERGILLUS CARBONARIUS S-CSR-0007

2016 ◽  
Vol 8 (12) ◽  
pp. 142 ◽  
Author(s):  
Afini A.v. M. ◽  
Sooraj S. Nath ◽  
Smitha K. V. ◽  
Kunhi A.a. M.
Keyword(s):  
Ammonium Sulfate ◽  
Substrate Concentration ◽  
Fibrinolytic Activity ◽  
Blood Clot ◽  
Maximum Activity ◽  
Fibrinolytic Enzyme ◽  
Clot Lysis ◽  
Aspergillus Carbonarius ◽  
Lysis Activity ◽  
Wide Range

<p><strong>Objective: </strong>This work was undertaken with the aim of isolating and screening fungal soil isolates with fibrinolytic activity.</p><p><strong>Methods: </strong>Soil sample near slaughter house was collected and screened for fibrinolytic activity by using fibrin-agar. Enzyme production was optimized under various parameters like pH, temperature, substrate concentration and purified partially by ammonium sulphate precipitation. The stability of the partially purified enzyme was analyzed under the influence of a wide range of pH, temperature, and substrate concentrations.<strong></strong></p><p><strong>Results: </strong>Among the seven isolates screened, <em>Aspergillus carbonarius</em> S-CSR-0007 exhibited largest clear zone and was selected for further studies. Among the various substrates tested casein was found to support the highest caseinolytic activity of 816 U/ml and fibrinolytic activity of 510 U/ml. The culture supernatant of <em>A. carbonarius</em> S-CSR-0007 was fractionated by ammonium sulfate precipitation followed by dialysis, and maximum activity was obtained in the fraction with 80% ammonium sulfate, with an enzyme activity of 1200 U/ml using tyrosine as standard. The partially purified fibrinolytic enzyme showed optimal activity at 45 °C and pH 7.0. The enzyme was stable up to a temperature of 50 °C and pH 8.0, and the optimum substrate concentration was 4%.</p><p><strong>Conclusion: </strong>The crude enzyme showed high blood clot lysis activity, which may be a good candidate in the pharmaceutical industry. However, more studies need to be carried out to establish its clinical use.</p>

scholarly journals Novel Sequential Screening and Enhanced Production of Fibrinolytic Enzyme byBacillussp. IND12 Using Response Surface Methodology in Solid-State Fermentation

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Ponnuswamy Vijayaraghavan ◽  
P. Rajendran ◽  
Samuel Gnana Prakash Vincent ◽  
Arumugaperumal Arun ◽  
Naif Abdullah Al-Dhabi ◽  
...  
Keyword(s):  
Response Surface Methodology ◽  
Solid State ◽  
Response Surface ◽  
Solid State Fermentation ◽  
Enzyme Production ◽  
Blood Clot ◽  
Fibrinolytic Enzyme ◽  
Surface Model ◽  
Cow Dung ◽  
Enhanced Production

Fibrinolytic enzymes have wide applications in clinical and waste treatment. Bacterial isolates were screened for fibrinolytic enzyme producing ability by skimmed milk agar plate using bromocresol green dye, fibrin plate method, zymography analysis, and goat blood clot lysis. After these sequential screenings,Bacillussp. IND12 was selected for fibrinolytic enzyme production.Bacillussp. IND12 effectively used cow dung for its growth and enzyme production (687±6.5 U/g substrate). Further, the optimum bioprocess parameters were found out for maximum fibrinolytic enzyme production using cow dung as a low cost substrate under solid-state fermentation. Two-level full-factorial experiments revealed that moisture, pH, sucrose, peptone, and MgSO4were the vital parameters with statistical significance (p<0.001). Three factors (moisture, sucrose, and MgSO4) were further studied through experiments of central composite rotational design and response surface methodology. Enzyme production of optimized medium showed4143±12.31 U/g material, which was more than fourfold the initial enzyme production (978±36.4 U/g). The analysis of variance showed that the developed response surface model was highly significant (p<0.001). The fibrinolytic enzyme digested goat blood clot (100%), chicken skin (83±3.6%), egg white (100%), and bovine serum albumin (29±4.9%).

Biosynthesis and characterization of a novel fibrinolytic alkaline serine protease from newly isolated Bacillus flexusBF12 for biomedical applications

2020 ◽  
Vol 21 ◽  
Author(s):  
T. Sterlin Raj ◽  
S. Athimoolam ◽  
P. Vijayaraghavan
Keyword(s):  
Enzyme Activity ◽  
Enzyme Production ◽  
Blood Clot ◽  
Fibrinolytic Enzyme ◽  
Physical Factors ◽  
Alkaline Serine Protease ◽  
Fibrinolytic Enzymes ◽  
Nutritional Factors ◽  
Thrombolytic Agents ◽  
Bacillus Flexus

Background: Cardiovascular diseases (CVDs) such as stroke, high blood pressure, peripheral vascular disease, ischemic heart disease and acute myocardial infarction are some of the leading causes of death. To treat CVDs, commercially available thrombolytic agents are widely used. However, these thrombolytic agents have various side effects. Alternatively, fibrinolytic enzymes from bacterial sources are highly safe and have direct blood clot lytic activity. Methods: A fibrinolytic enzyme producing bacterial strain, Bacillus flexus BF12, was isolated from a solar saltpan in Kanyakumari District, Tamilnadu, India. Enzyme production was improved by optimizing physical factors and nutritional factors. Results: A novel fibrinolytic enzyme was isolated from a strain of the studied B. flexus BF12. Enzyme production was enhanced significantly by optimizing process parameters. The critical physical factors (pH and salinity) and influencing nutritional factors (carbon, nitrogen and ions) were optimized by one variable at a time approach, followed by statistical method. The strain BF12 was highly active at alkaline pH (>7.0) and between 4 and 6% NaCl concentration. The nutrients such as fructose (carbon source), beef extract (nitrogen source) and CaCl2 significantly influenced enzyme production. Central composite design and response surface methodology improved 3.2-fold enzyme yield than unoptimized culture medium. Fibrinolytic protease was purified by ammonium sulphate precipitation, dialysis and gel filtration chromatography. Discussion: The molecular weight of an enzyme was found to be 23 kDa. It was active at a broad temperature (40-60 °C) and pH (7.0-9.0) ranges. Enzyme activity was enhanced by Ca2+ and Co2+ ions. The purified protease retained 100% enzyme activity in the presence of ethanol and acetone. Acetonitrile, butanol, DMSO, methanol and chloroform showed enzyme activity of 63%, 92.5%, 94.7%, 92.3% and 90.4%, respectively. The purified enzyme degraded 100% of human blood clot. Conclusion: The Bacillus flexus BF12 fibrinolytic enzyme shows promising potentials in nutraceutical and food fortification applications. The application of fibrinolytic enzymes could prevent CVDs.

Food sources of phyto-oestrogens and their precursors in Europe

2003 ◽  
Vol 89 (S1) ◽  
pp. S39-S43 ◽  
Author(s):  
R. J. Fletcher
Keyword(s):  
Meat Products ◽  
Soya Protein ◽  
Raw Material ◽  
Food Sources ◽  
Fermented Foods ◽  
Wide Range ◽  
Cereal Brans ◽  
Final Water Content ◽  
Growing Conditions

Phyto-oestrogens are dietary components found in some plants, which actin vivolike weak oestrogens. They may reduce the risk of some degenerative diseases moderated by oestrogen, including breast cancer and osteoporosis. The most widely studied are the isoflavones genistein and daidzein from soyabeans, but lignans may be more prevalent in the European diet. Soya foods have traditionally been consumed in the Orient for millennia, and are now widely available to European consumers. Levels of isoflavone in soyabeans from published literature vary between 560 and 3810 mg/kg, depending on variety and growing conditions. Soya protein concentrates and isolates derived from soyabeans contain 466–615 mg isoflavones/kg. Traditional soya milk, bean curds, bean sprouts, etc. contain 13 to 2030 mg isoflavone/kg, depending on the starting raw material and final water content. Fermented foods have a different isoflavone conjugate profile, which may be important in absorption and metabolism. Soya analogues of European foods include dairy and meat products, which contain 38 to 3000 mg total isoflavones/kg, depending the source of soya and dilution with other ingredients. A wide range of foods contain low levels of soya-derived isoflavones, but such foods do not make a significant contribution to mean intakes in Europe. Flaxseed is by far the richest source of lignan precursors. However, foods such as cereal brans, legumes and some vegetables are a more important source in the diets of Europeans because they are more widely consumed. For similar reasons, compared with soya isoflavones, lignans may be a more important source of phyto-oestrogens in the diets of Europeans.

scholarly journals Probiotic Attributes, GABA-Producing Capacity and Cytotoxic Effects of Microbes Isolated from Thai Fermented Foods

2021 ◽  
Vol 25 (02) ◽  
pp. 409-419
Author(s):  
Vijitra Luang-In
Keyword(s):  
Monosodium Glutamate ◽  
Fermented Food ◽  
Mental Illnesses ◽  
Food Sources ◽  
Fermented Foods ◽  
Gamma Aminobutyric Acid ◽  
Bifidobacterium Adolescentis ◽  
Cytotoxic Effects ◽  
Bacillus Spp ◽  
Mcf 7

Mental illnesses and cancer deaths have been alarmingly increased in Thailand over the past decade. There is a search for an alternative remedy from natural sources. This work revealed the probiotic attributes of eighteen microbes from Thai fermented food sources, their neurotransmitter-producing capacities and cytotoxic effects. The probiotic attributes were comprehensively assessed by gelatin hydrolysis, antibiotic susceptibility, auto-aggregation, virulence gene detection, hemolytic activity, hydrophobicity determination, bile and acid tolerance. Neurotransmitters were produced from precursors by microbes over 3 days and analyzed by high-throughput LC-MS/MS. Cytotoxic effects of the selected microbial extracts on MCF-7, HepG2, and HeLa were determined using MTT assay. The results showed that only seven strains; Enterococcus casseliflavus 3.10A1, Enterobacter xiangfangensis 4A-2A3.1, Saccharomyces cerevisiae TC6, Pedicoccus pentosaceus WS11, Lactococcus lactis subspp. lactis TBRC 375, Lactobacillus brevis TBRC 3003 and Bifidobacterium adolescentis TBRC 7154 exhibited promising probiotic potential in every probiotic assessment. Only E. xiangfangensis 4A-2A3.1 and Bacillus spp. PS15 were capable of producing gamma-aminobutyric acid (GABA) from monosodium glutamate (20 mg/mL). GABA production of 4.60 μg/mL over 2 days was found in E. xiangfangensis 4A-2A3.1; however, Bacillus spp. PS15 produced the highest GABA at 5.57 μg/mL on day 1. Moreover, B. adolescentis TBRC 7154 extract showed the lowest IC50 values against MCF-7 (750.02 μg/mL), HepG2 (681.08 μg/mL), and HeLa (425.50 μg/mL) followed by E. xiangfangensis 4A-2A3.1, E. casseliflavus 3.10A1 and P. pentosaceus WS11. These microbes can be used as probiotic supplement with neurodegeneration preventive or chemopreventive potential. © 2021 Friends Science Publishers

scholarly journals ISOLATION AND CHARACTERIZATION OF ACTIVE PROBIOTIC ORGANISMS IN FERMENTED ZEA MAYS, PENTADETHRA MACROPHYLLA BENTH,AND RICINUS COMMUNIS

2021 ◽  
Vol 9 (4) ◽  
pp. 512-516
Author(s):  
Sunday S. Agbem ◽  
Keyword(s):  
Ricinus Communis ◽  
Food Samples ◽  
Primary Objective ◽  
Fermented Food ◽  
Bacillus Species ◽  
Fermented Foods ◽  
Isolation And Characterization ◽  
Micro Organisms ◽  
Protein Rich

Active probiotic organisms are good bacteria considered to be live micro-organisms that are obtained from fermented foods. Evidence has shown that probiotics are essential in human health. This studys primary objective is to isolate and characterize the active probiotic organisms present in certain fermented food samples. Maize, African oil bean, and castor oil were subjected to analysis. The result found the presence of active probiotic organisms such as Pedioccocus, Lactobacillus, Micrococcus, and Bacillus species. The study concluded that these organisms are responsible for the fermentation of carbohydrates and protein-rich seeds.

scholarly journals The Antioxidant, Anti-Diabetic, and Anti-Adipogenesis Potential and Probiotic Properties of Lactic Acid Bacteria Isolated from Human and Fermented Foods

Fermentation ◽  
2021 ◽  
Vol 7 (3) ◽  
pp. 123
Author(s):  
Yulah Jeong ◽  
Hyemin Kim ◽  
Ji Yeon Lee ◽  
GaYeong Won ◽  
Soo-Im Choi ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Radical Scavenging ◽  
Fermented Food ◽  
Food Sources ◽  
Fermented Foods ◽  
Probiotic Properties ◽  
Beneficial Effects ◽  
Safety And Stability ◽  
Oil Red O Staining

In this study, lactic acid bacteria (LAB) strains derived from human and fermented food sources were examined to identify their properties related to obesity, as well as establish their safety and stability as probiotics. LAB (Lacticaseibacillus rhamnosus MG4502, Lactobacillus gasseri MG4524, Limosilactobacillus reuteri MG5149, and Weissella cibaria MG5285) exhibited antioxidant activity through DPPH (>26.1%) and ABTS (>40.1%) radical scavenging assays and α-glucosidase inhibitory activities (>60.3%), respectively. The LAB strains promoted anti-adipogenesis by reducing lipid accumulation in 3T3-L1 cells by Oil Red O staining (>70.3%). In addition, we found that these LAB strains were resistant to simulated gastric and intestinal fluids (pH 3, 4, 7, and 8) and showed potential for health promotion, based on hemolysis, cell adhesion, antibiotic susceptibility, and enzyme production. Thus, LAB may be used as probiotic ingredients with beneficial effects.

Hominoid Adaptation to Dietary Ethanol

2019 ◽  
pp. 24-44
Author(s):  
Matthew Carrigan
Keyword(s):  
Blood Alcohol Concentration ◽  
Alcohol Concentration ◽  
Fermented Food ◽  
Ethanol Metabolism ◽  
Primate Species ◽  
Fermented Foods ◽  
Wide Range ◽  
Fallback Food ◽  
Biochemical Analyses ◽  
Special Case

Humans’ long association with alcohol raises questions about both our biological adaptations to handling ethanol and its origins. Fermented foods have less sugar, and require additional detoxification than unfermented versions of the same food, and are thus are generally inferior food choices. I summarize recent studies which indicate that our ability to exploit ethanol depends on several mutations in alcohol dehydrogenase (ADH) enzymes that allow ethanol to be metabolized rapidly, thereby reducing the likelihood that the blood alcohol concentration reaches intoxicating levels. Genetic and biochemical analyses for a wide range of primate and non-primate species suggest that these mutations are shared primarily with the two African great apes (chimpanzees and gorillas). These mutations thus date back at least 10 million years, to a period when the tropical forests were contracting during a major episode of climate change. Mutations enabling rapid ethanol metabolism may have enabled ancestral apes to exploit otherwise toxic, ethanol-rich fermenting fruits on the forest floor that were metabolically inaccessible to their ecological competitors. These adaptations enabling exploitation of an inferior food suggest that modern proclivities towards ethanol consumption may derive from the utilization of fermented food as a particular type of fallback food. If so, the fermented fallback food hypothesis can be seen as a special case of the drunken monkey hypothesis.

scholarly journals Therapeutic Potential of Quercetin to Alleviate Endothelial Dysfunction in Age-Related Cardiovascular Diseases

2021 ◽  
Vol 8 ◽  
Author(s):  
Olina Dagher ◽  
Pauline Mury ◽  
Nathalie Thorin-Trescases ◽  
Pierre Emmanuel Noly ◽  
Eric Thorin ◽  
...  
Keyword(s):  
Cardiovascular Diseases ◽  
Endothelial Dysfunction ◽  
Therapeutic Potential ◽  
Scientific Evidence ◽  
Epidemiological Studies ◽  
Food Sources ◽  
Physically Active ◽  
Mesenchymal Transition ◽  
Age Related ◽  
Wide Range

The vascular endothelium occupies a catalog of functions that contribute to the homeostasis of the cardiovascular system. It is a physically active barrier between circulating blood and tissue, a regulator of the vascular tone, a biochemical processor and a modulator of coagulation, inflammation, and immunity. Given these essential roles, it comes to no surprise that endothelial dysfunction is prodromal to chronic age-related diseases of the heart and arteries, globally termed cardiovascular diseases (CVD). An example would be ischemic heart disease (IHD), which is the main cause of death from CVD. We have made phenomenal advances in treating CVD, but the aging endothelium, as it senesces, always seems to out-run the benefits of medical and surgical therapies. Remarkably, many epidemiological studies have detected a correlation between a flavonoid-rich diet and a lower incidence of mortality from CVD. Quercetin, a member of the flavonoid class, is a natural compound ubiquitously found in various food sources such as fruits, vegetables, seeds, nuts, and wine. It has been reported to have a wide range of health promoting effects and has gained significant attention over the years. A growing body of evidence suggests quercetin could lower the risk of IHD by mitigating endothelial dysfunction and its risk factors, such as hypertension, atherosclerosis, accumulation of senescent endothelial cells, and endothelial-mesenchymal transition (EndoMT). In this review, we will explore these pathophysiological cascades and their interrelation with endothelial dysfunction. We will then present the scientific evidence to quercetin's anti-atherosclerotic, anti-hypertensive, senolytic, and anti-EndoMT effects. Finally, we will discuss the prospect for its clinical use in alleviating myocardial ischemic injuries in IHD.

scholarly journals Optimization of Cultural Conditions for Maximum Production of Fibrinolytic Enzymes from the Local Marine Bacterial Isolates and Evaluation of their Wound Healing and Clot Dissolving Properties

2021 ◽  
pp. 246-255
Author(s):  
K. Gowthami ◽  
R. Jaya Madhuri
Keyword(s):  
Wound Healing ◽  
Enzyme Activity ◽  
Incubation Time ◽  
Specific Activity ◽  
Blood Clot ◽  
Inoculum Size ◽  
Fibrinolytic Enzyme ◽  
Physical Parameters ◽  
Bacterial Strains ◽  
Fibrinolytic Enzymes

Fibrinolytic enzymes find necessary applications to treat and prevent cardiovascular diseases.  In this study, optimal conditions for enhancing the production of fibrinolytic enzyme from local marine bacterial strains were evaluated. The present study also focuses on screening of wound                  healing efficacy of the isolated fibrinolytic enzymes.Various physical parameters such as temperature, pH, incubation time and medium components viz. inoculum size, substrate (nitrogen and carbon) concentrations were optimized. A cultivation medium was designed using optimized conditions for mass production of fibrinolytic enzyme and specific activity of enzyme was analyzed. The maximum enzyme production was observed at 37 °C temperature, 8.0 pH,substrate concentration with 3 ml inoculum size and 32 h. of incubation time. Among the different carbon sources tested, Mannitol showed maximum enzyme activity i.e 538 U/ml.  yeast extract was found to be the best nitrogen source with an enzyme activity of 498 U/ml.  The best substrate for the production fibrinolytic enzyme was found to be kernelwith high  activity of 1056U/ml. The crude enzyme displayed potent activity and digested blood clot completely in in vitro condition and exhibited potent activity on wound healing property in macrophages.

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