scholarly journals Antimicrobial properties of the peptide substance Bacillus Subtilis PSF-19

2019 ◽  
pp. 22-26
Author(s):  
Viktor Danilovich Pokhilenko ◽  
Vladimir Vladimirovich Perelygin ◽  
Timur Akhmerovich Kalmantaev ◽  
Konstantin Vladimirovich Detushev ◽  
Irina Anatolevna Chukina
Keyword(s):  
Molecular Weight ◽  
Bacillus Subtilis ◽  
Pathogenic Bacteria ◽  
Proteolytic Enzymes ◽  
Raw Materials ◽  
Antimicrobial Properties ◽  
Culture Fluid ◽  
Fluid Fraction ◽  
Intestinal Pathogens ◽  
Vegetable Raw Materials

The subject of the study is a strain of Bacillus subtilis PSF-19 isolated by us from vegetable raw materials (Passiflora preparation), which is capable of producing antimicrobial substance (AMV) suppressing pathogenic bacteria. The article discusses the method and conditions of extraction from the culture fluid fraction of AMV, active against Listeria monocytogenes – one of the dangerous intestinal pathogens that infect food. Using biochemical methods and mass spectroscopy, the molecular weight and the peptide nature of the active fraction of AMV were determined. A fraction of AMV with a molecular weight of 3,4–3,6 kDa has bactericidal activity, which is destroyed by treatment with proteolytic enzymes, which allowed it to be attributed to the group of low-molecular antimicrobial peptides – bacteriocins. The studies allow to consider the strain Bacillus subtilis PSF-19 as a producer of bacteriocin, especially effective against pathogens of intestinal listeriosis. The establishment of the fact of destructibility of proteolytic enzymes, finding the conditions of microbiological synthesis of AMV, its isolation and accumulation for research, provide opportunities for practical use as a means for decontamination of the environment instead of traditional antibiotics.

Technology of production of biopesticides based on Bacillus thuringiensis

2021 ◽  
Vol 9 (16) ◽  
pp. 28-38
Author(s):  
Anatolii Bezusov ◽  
◽  
Valentyna Krutiakova ◽  
Olena Myroshnichenko ◽  
Nataliia Dotsenko ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Corn Oil ◽  
Raw Materials ◽  
Culture Fluid ◽  
Scientific Work ◽  
Insect Larvae ◽  
Effective Prevention ◽  
Research Results ◽  
Wide Range ◽  
Vegetable Raw Materials

Subject of research. Biopesticides are based on live cultures of specially selected beneficial microorganisms with controlled properties. They have a pronounced phytoprotective and stimulating effect, thus providing effective prevention and protection of plants from diseases. The obtained biological product increases productivity, improves the quality and structure of the crop, does not accumulate in plants, which allows to obtain environmentally friendly agricultural products and does not harm the environment. Among the large number of bacteria, as a source of microbiological insecticide is Bacillus thuringiensis, which infects lepidopteran pests and leads to their death. Preparations based on this strain are effective when used in low concentrations of solutions. The Bacillus thuringiensis strain produces several toxins with insecticidal action, including β-exotoxin and δ-endotoxin. Toxic effect is manifested and leads to paralysis of the intestinal tract of parasites. Preparations β-exotoxin and δ-endotoxin are obtained by culturing Bacillus thuringiensis bacteria in a liquid medium. The scientific work proposes a method of industrial production using by-products of vegetable raw materials, which makes it economically feasible to use such substrates. The purpose of the study is the development of technology, formulation of nutrient medium, process parameters for the cultivation of bacteria of the genus Bacillus thuringiensis and obtaining a culture fluid containing substances of the class of biopesticides. Methods. Standard and generally accepted methods of research of bioproducts in biotechnology. The formation of bioinsecticides was established by hydrolysis methods, followed by determination of the component of β-exotoxin – ribose, the formation of octagonal crystals of exotoxin – by microscopic method. Research results. Three variants of nutrient media, which include yeast-polysaccharide complex: corn flour, corn oil, yeast autolysate were developed. The parameters of the bacterial cultivation process were studied. The final product is a paste or powder with a titer of 35 .109 spores in 1 g of the bioproduct. Scope of research results. Microbiological preparations based on Bacillus thuringiensis are highly specific and act only on insect larvae from the classes Lepidoptera and Diptera. The resulting biopesticide can be used against pests of a wide range of vegetable and fruit crops.

scholarly journals Antibacterial Activity, Cytotoxicity, and the Mechanism of Action of Bacteriocin from Bacillus subtilis GAS101

2018 ◽  
Vol 27 (2) ◽  
pp. 186-192 ◽  
Author(s):  
Garima Sharma ◽  
Shweta Dang ◽  
Sanjay Gupta ◽  
Reema Gabrani
Keyword(s):  
Molecular Weight ◽  
Antibacterial Activity ◽  
Bacillus Subtilis ◽  
Vero Cell ◽  
Cell Line ◽  
Mode Of Action ◽  
Proteolytic Enzymes ◽  
Antibiofilm Activity ◽  
Vero Cell Line ◽  
Soil Isolate

Objective: The aim of this study was to purify and characterize bacteriocin from the soil isolate Bacillus subtilis GAS101, and to determine its antimicrobial as well as antibiofilm potential. The purified bacteriocin was further analyzed and evaluated for mammalian cell cytotoxicity and the possible mode of action. Material and Methods: Bacteriocin from B. subtilis GAS101 (an animal husbandry soil isolate) was partially purified and checked for antimicrobial and antibiofilm activity against gram-positive and gram-negative bacteria. The molecular weight of bacteriocin was determined using tricine SDS-PAGE gel. The stability of bacteriocin was investigated at various temperatures and pH levels, and its sensitivity towards 8 enzymes and 6 chemicals was determined. Cytotoxicity analysis was performed on a Vero cell line by a tetrazolium dye-based assay. Scanning electron microscopy (SEM) of bacteriocin-treated bacteria was carried out to determine the possible mode of action. Results: Bacteriocin from B. subtilis GAS101 was a potential inhibitor of both the indicator organisms (Staphylococcus epidermidis and Escherichia coli), and had a molecular weight of approximately 6.5 kDa. An in situ gel assay showed a zone of inhibition corresponding to the estimated protein band size. Bacteriocin was stable and showed antibacterial activity in broad ranges of temperature (30–121°C) and pH (2–12). It was sensitive to 4 proteolytic enzymes, which indicated its proteinaceous nature. Bacteriocin showed > 70% cell viability on the mammalian Vero cell line. SEM depicted that the bacteriocin was able to disrupt the bacterial cell membrane as its probable mode of action. Conclusion: Thermostable and pH-tolerant bacteriocin from B. subtilis GAS101, of about 6.5 kDa, showed broad-spectrum antimicrobial and antibiofilm activity.

scholarly journals ASSESSMENT ON ANTIMICROBIAL PROPERTIES OF SURFACTIN FROM BACILLUS SUBTILIS ON PROTOPLASTS AND SPHEROPLASTS OF PATHOGENIC BACTERIA

2016 ◽  
Vol 9 (1) ◽  
pp. 67
Author(s):  
B. Chitra ◽  
A. Benno Susai Vijayakumar
Keyword(s):  
Bacillus Subtilis ◽  
Pathogenic Bacteria ◽  
Inhibitory Concentration ◽  
Antimicrobial Properties ◽  
Polluted Soil ◽  
Antibiotic Activity ◽  
Lytic Activity ◽  
Haemolytic Activity ◽  
Pathogenic Microorganisms ◽  
Clear Zone

Objective: The aim of this paper is to assessment on antimicrobial properties of surfactin from Bacillus subtilis on protoplasts and spheroplasts of pathogenic bacteria.Methods: Bacillus sp were isolated from the polluted soil samples and these isolates were screened for biosurfactin activity using haemolytic activity. The biosurfactant was also studied for their antibiotic activity and the minimum inhibitory concentration was determined by testing the effect of surfactin on the pathogenic microorganisms. The lytic activity of surfactant was also tested on the protoplasts and spheroplasts of the pathogens.Results: The formation of a clear zone around the colonies and the lysis of erythrocytes shows the antimicrobial and lytic activity of biosurfactants.Conclusion: The results of the current study reveal that surfactin from B. subtilis possesses significant antimicrobial and lytic activity.

scholarly journals Bacteriocins of lactic acid bacteria and their application on food as biopreservatives. (II)

2017 ◽  
Vol 54 (1) ◽  
pp. 69
Author(s):  
J. METAXOPOULOS (Ι. ΜΕΤΑΞΟΠΟΥΛΟΣ) ◽  
M. MATARAGAS (M. ΜΑΤΑΡΑΓΚΑΣ) ◽  
E. H. DROSINOS (Ε.Χ. ΔΡΟΣΙΝΟΣ)
Keyword(s):  
Carbon Dioxide ◽  
Hydrogen Peroxide ◽  
Molecular Weight ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
Antimicrobial Properties ◽  
Small Molecular Weight ◽  
Inhibitory Spectrum ◽  
Protective Cultures

Lactic acid bacteria produce a variety of small molecular weight compounds, which have antimicrobial properties. Such substances are: organic acids, alcohols, carbon dioxide, diacetyl, hydrogen peroxide and bacteriocins. Many of these compounds have a wide inhibitory spectrum but the bacteriocins are able to inhibit species, namely, related with the bacteriocin-producing strain. In the last years bacteriocins have gained a lot of concern because some of them are able to inhibit the growth of pathogenic bacteria, like Listeria monocytogenes. The term "biopreservation" refers to the extension of storage life, as well as to the enhancement of the food safety, using the bacteriocin-producing lactic acid strains or their metabolic antibacterial products. In this review will be reported bacteriocins, which are produced by the lactic acid bacteria and will be discussed the potential application of the bacteriocinogenic strains or their bacteriocins on the foods, as protective cultures or as protective compounds, respectively.

Physical-Chemical and Cytotoxic Properties of Bacillus subtilis IMV B-7724 Extracellular Lectin

2021 ◽  
Vol 83 (1) ◽  
pp. 39-48
Author(s):  
N.I. Fedosova ◽  
◽  
N.L. Cheremshenko ◽  
K.I. Hetman ◽  
T.V. Symchych ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Bacillus Subtilis ◽  
Elemental Composition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Culture Fluid ◽  
Biological Properties ◽  
Composition Analysis ◽  
Clinical Settings ◽  
Cultural Methods ◽  
Student’S T

Aim. To study the chemical composition, sugar specificity and physicochemical properties of the extracellular lectin isolated from Bacillus subtilis ІМV В-7724. Methods. Biochemical, spectrophotometric, immunological and cultural methods were used to assess the physicochemical and a number of biological properties of lectin isolated from the culture fluid of bacteria B. subtilis ІМV В-7724. Molecular weight of the lectin was estimated in polyacrylamide gel electrophoresis. Analysis of the elemental composition was done using Perkin-Elmer 2400 CHNS analyzer. Temperature and pH stability of lectin were examined based on residual hemagglutination activity of the lectin. Cytotoxic activity was determined by the MTT-assay. The statistical analysis was made using Student’s t-test. Results. B. subtilis IMV B-7724 lectin is a glycoprotein (protein – 86.0%, carbohydrates – 7.0%) with molecular weight of 18–20 kDa (major). Analysis of the elemental composition revealed that it contains 34.00% of carbon, 7.04% of hydrogen, 16.61% of nitrogen, 42.35% of oxygen. Amino acid composition analysis determined that it is rich in leucine, tyrosine and phenylalanine. The lectin exhibited high sugar-binding specificity toward N-acetylneuraminic and N-glycolylneuraminic acids (minimal inhibitory concentration – 0.3 mM for both sugars). The lectin is heat and acid stable, has long shelf life. Conclusions. These results provide the rationale to pursue further investigation for possible ways and modes of B. subtilis IMB B-7724 lectin application in clinical settings.

Physico-chemical and cytotoxic analysis of a novel large molecular weight bacteriocin produced by Lactobacillus casei TA0021

2019 ◽  
Author(s):  
Elham Noroozi ◽  
Naheed Mojgani ◽  
Elahe Motevaseli ◽  
Mohammad Hossein Modarressi ◽  
Majid Tebianian
Keyword(s):  
Molecular Weight ◽  
Lactobacillus Casei ◽  
Pathogenic Bacteria ◽  
Proteolytic Enzymes ◽  
Shigella Flexneri ◽  
Gel Filtration ◽  
Antimicrobial Protein ◽  
Purification Method ◽  
Ph Values ◽  
Subsequent Decrease

Background and Objectives: Antimicrobial peptides produced by lactic acid bacteria have gained enormous attention owing to their health benefits. This study aimed to isolate, purify and characterize the antibacterial protein produced by au- tochthonous Lactobacillus casei TA0021 strain. Materials and Methods: The antagonistic activity of L. casei TA0021 against a number of pathogenic bacteria was tested by agar well diffusion assay. The antimicrobial agent in the neutralized supernatant fluids was subjected to the action of proteolytic enzymes, catalase, lipase and lysozyme, and their tolerance to variable pH and temperature was estimated. The proteinaceous antagonistic compound was precipitated by 60% w/v ammonium sulphate, desalted and subjected to cation exchange and gel filtration chromatography. Approximate molecular weight of Lactocin was determined by SDS-PAGE and non-denaturing gel electrophoresis. Hemoglobin release assay and cytotoxicity effect of Lactocin TA0021 was determined. The results were statistically analyzed. Results: The antagonistic agent active against Salmonella Typhimurium and Shigella flexneri appeared resistant to catalase and lipase treatments, while sensitive to the tested proteolytic enzymes. Lactocin TA0021 resisted acidic pH values of 3.0, while alkaline pH values of ˃9 completely destroyed the activity. The antibacterial peptide was approximately 68 KDa and heat labile as lost its activity at 100°C after 5 minutes. The bacteriocin was non-toxic to MRC-5 cell lines and non-hemolytic. Purification method lead to increase in antibacterial activity while, subsequent decrease in recovery and yield was observed with increasing purification fold. Conclusion: The purified antimicrobial protein from L. casei TA0021 might be used for application in medicinal and food products.

scholarly journals STUDY OF ANTIMICROBIAL ACTIVITY OF THE FOLKLORE MEDICINAL PLANT, ACALYPHA FRUTICOSA FORSSK.

2017 ◽  
Vol 4 (2) ◽  
pp. 57-60
Author(s):  
Thambiraj J
Keyword(s):  
Antimicrobial Activity ◽  
Bacillus Subtilis ◽  
Medicinal Plant ◽  
Ethyl Acetate ◽  
Pathogenic Bacteria ◽  
Antimicrobial Properties ◽  
Pathogenic Fungi ◽  
Inhibition Zone ◽  
Diffusion Method ◽  
Rhizopus Sp

The purpose of the study is to examine the antimicrobial efficacy of root extracts of the folklore medicinal plant species, Acalypha fruticosa Forssk by using three alcoholic solvents viz; petroleum ether, ethyl acetate and methanol were tested against ten human pathogenic bacteria viz., Pseudomonas aeruginosa, P. stutzeri, Escherichia coli, Micrococcus sp., Lactobacillus sp., Servatia sp., Moraxetta sp., Bacillus subtilis, B. thuriengensis, and Klebsiella pneumoniae and ten human pathogenic fungi viz., Aspergillus niger, A. flavus, A.baumannii, Fusarium oxysporum, F. solani, Mucor rouxii, Alternaria alternata, Candida albicans, Cladosporium sp. and Rhizopus sp. for assessing the antimicrobial properties by adapting disc diffusion method. The results of the study revealed that all extracts showed varied degree of antimicrobial activity against the tested pathogens. However, the methanol extracts exhibited higher inhibition zone (21.83 mm) against the bacterium, Bacillus subtilis and ethyl acetate extracts showed higher inhibiotion zone (24.83 mm) against the fungus, Rhizopus sp. Results concluded that this spices contain high amount of secondary metabolites due to these metabolites they have high antimicrobial activity and it can be used as good bio- preservater and it can also use for medicinal purpose.

scholarly journals ANTIMICROBIAL EFFICACY OF THE FOLKLORE MEDICINAL PLANT, ACACIA CAESIA (L.) WILD

2015 ◽  
Vol 2 (2) ◽  
pp. 110-113
Author(s):  
Thambiraj J ◽  
Paulsamy S
Keyword(s):  
Bacillus Subtilis ◽  
Ethyl Acetate ◽  
Pathogenic Bacteria ◽  
Antimicrobial Properties ◽  
Pathogenic Fungi ◽  
Inhibition Zone ◽  
Diffusion Method ◽  
Mucor Rouxii ◽  
Bacterium Bacillus ◽  
Rhizopus Sp

eaf extracts of the folklore plant species, Acacia caesia L. by using three alcoholic solvents viz; petroleum ether, ethyl acetate and methanol were tested against ten human pathogenic bacteria viz., Pseudomonas aeruginosa, P. stutzeri, Escherichia coli, Micrococcus sp., Lactobacillus sp., Servatia sp., Moraxetta sp., Bacillus subtilis, B. thuriengensis, and Klebsiella pneumoniae and ten human pathogenic fungi viz., Aspergillus niger, A. flavus, A.baumannii, Fusarium oxysporum, F. solani, Mucor rouxii, Alternaria alternata, Candida albicans, Cladosporium sp. and Rhizopus sp. for assessing the antimicrobial properties by adapting disc diffusion method. The results of the study revealed that all extracts showed varied degree of antimicrobial activity against the tested pathogens. However, the ethyl acetate extracts exhibited higher inhibition zone (15.73 mm) against the bacterium, Bacillus subtilis and the fungus, Mucor rouxii (20.67 mm). These results support the therapeutic importance of the species, Acacia caesia in curing infectious diseases and encourage the extensive use of this species in health care practices.

scholarly journals Application of plant proteolytic enzymes for tenderization of rabbit meat

2018 ◽  
Vol 34 (2) ◽  
pp. 229-238 ◽  
Author(s):  
Maria Doneva ◽  
Iliana Nacheva ◽  
Svetla Dyankova ◽  
Petya Metodieva ◽  
Daniela Miteva
Keyword(s):  
Proteolytic Enzymes ◽  
Raw Materials ◽  
Electrophoretic Pattern ◽  
Physicochemical Characteristics ◽  
Water Holding Capacity ◽  
Rabbit Meat ◽  
Vegetable Raw Materials ◽  
Meat Samples ◽  
Water Holding ◽  
Control Samples

The purpose of this study is to assess the tenderizing effect of plant proteolytic enzymes upon raw rabbit meat. Tests are performed on rabbit meat samples treated with papain and two vegetal sources of natural proteases (extracts of kiwifruit and ginger root). Two variants of marinade solutions are prepared from each vegetable raw materials- 50% (w/w) and 100 % (w/w), with a duration of processing 2h, 24h, and 48h. Changes in the following physicochemical characteristics of meat have been observed: pH, water-holding capacity, cooking losses and quantity of free amino acids. Differences in values of these characteristics have been observed, both between control and test samples, as well as depending of treatment duration. For meat samples marinated with papain and ginger extracts, the water-holding capacity reached to 6.74 ? 0.04 % (papain), 5.58 ? 0.09 % (variant 1) and 6.80 ? 0.11 % (variant 2) after 48 hours treatment. In rabbit meat marinated with kiwifruit extracts, a significant increase in WHC was observed at 48 hours, 3.37 ? 0.07 (variant 3) and 6.84 ? 0.11 (variant 4). The test samples also have reduced cooking losses compared to control samples. In control samples, cooking loss is increased from 13.79% (2 h) to 20.78 % (48 h). SDS-PAGE of meat samples after 48 h of treatment shows a reduction in the intensity of actin and myosin bands in all variants with papain and vegetal extracts. Electrophoretic pattern of test samples depicts proteolysis and degradation of muscle proteins.

scholarly journals Technology and Physico-chemical Properties of Gelatin from Atlantic Cod Skin

2020 ◽  
Author(s):  
Daria Kolotova ◽  
Lyudmila Petrova
Keyword(s):  
Molecular Weight ◽  
Barents Sea ◽  
Atlantic Cod ◽  
Proteolytic Enzymes ◽  
Raw Materials ◽  
Chemical Properties ◽  
Fish Gelatin ◽  
Peptide Bonds ◽  
Physico Chemical

In this work, the technology of fish gelatin from the skin of Atlantic cod from the Barents Sea was developed. The influence of enzyme type, extraction time, and pH of extraction on properties of fish gelatin was studied. The usage of proteolytic enzymes increases the yield of gelatin in comparison with acidic treatment, but the molecular weight of gelatin and viscosity of its solution decrease. The results observed related to the cleavage of peptide bonds of collagen during the enzymolysis process. The increase the time of extraction leads to an increase in molecular weight and quality of gelatin obtained. It was shown that gelatin obtained can be successfully used in the production of functional food products based on fish raw materials. The results obtained are of great importance for the fish-processing industry. They allow transforming existing technologies into non-waste processes and thereby enhance the effectiveness of enterprises and create better environmental conditions.

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