Characterization of a Potent New-Generation Antimicrobial Peptide of Bacillus

An antimicrobial peptide [Bacillus antimicrobial peptide (BAMP)] produced by Bacillus paralicheniformis was isolated from the Indian traditional fermented food and characterized. The antimicrobial peptide BAMP showed many unique features such as thermostability (4.0–125°C), pH tolerance (pH 2.0–9.0), and resistance to physiological enzymes (trypsin, chymotrypsin, pepsin, proteinase K, protease, and catalase), and food-grade metal salts do not inhibit the activity. The broad spectrum of BAMP (antimicrobial activity) makes it a suitable candidate for food preservation as well as antimicrobial therapy. BAMP was found to exhibit a bacteriostatic effect on Salmonella typhi and controls the viability of Listeria monocytogenes in chicken meat efficiently. BAMP was found to establish eubiosis, as it is not antagonistic to Lactobacillus. Its non-hemolytic nature makes it suitable for therapy. Various genome prediction tools were adopted and applied to understand their localization, gene arrangement, and type of antimicrobials. Founded on its superior functional attributes, BAMP is a potent new-generation antimicrobial peptide.

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In vitro and In Silico Approach For Characterization of Antimicrobial Peptide From Probiotics Against Staphylococcus Aureus and Escherichia Coli

10.21203/rs.3.rs-366314/v2 ◽

2021 ◽

Author(s):

Amrutha Bindu ◽

Lakshmi Devi

Keyword(s):

Amino Acid ◽

Antimicrobial Peptide ◽

Lactobacillus Plantarum ◽

Exchange Chromatography ◽

Amino Acid Sequences ◽

Proteinase K ◽

Kinetic Assay ◽

Wide Range

Abstract The focus of present study was to characterize antimicrobial peptide produced by probiotic cultures, Enterococcus durans DB-1aa (MCC4243), Lactobacillus plantarum Cu2-PM7 (MCC4246) and Lactobacillus fermentum Cu3-PM8 (MCC4233) against Staphylococus aureus and E. coli. The growth kinetic assay revealed 24 h of incubation to be optimum for bacteriocin production. The partially purified compound after ion-exchange chromatography was found to be thermoresistant and stable under wide range of pH. The compound was sensitive to proteinase-K, but resistant to trypsin, a-amylase and lipase. The apparent molecular weight of bacteriocin from MCC4243 and MCC4246 was found to be 3.5 KDa. Translated partial amino acid sequence of plnA gene in MCC4246 displayed 48 amino acid sequences showing 100% similarity with plantaricin A of Lactobacillus plantarum (WP_0036419). The sequence revealed 7 β sheets, 6 α sheets, 6 predicted coils and 9 predicted turns. The functions on cytoplasm show 10.82 isoelectric point and 48.6% hydrophobicity. The molecular approach of using Geneious Prime software and protein prediction data base for characterization of bacteriocin is novel and predicts “KSSAYSLQMGATAIKQVKKLFKKWGW” as peptide responsible for antimicrobial activity. The study provides information about broad spectrum bacteriocin in native probiotic culture and paves a way towards its application in functional foods as biopreservative agents.

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Plant-based Paste Fermented by Lactic Acid Bacteria and Yeast: Functional Analysis and Possibility of Application to Functional Foods

Biochemistry Insights ◽

10.4137/bci.s10529 ◽

2012 ◽

Vol 5 ◽

pp. BCI.S10529 ◽

Cited By ~ 8

Author(s):

Shinsuke Kuwaki ◽

Nobuyoshi Nakajima ◽

Hidehiko Tanaka ◽

Kohji Ishihara

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Pantothenic Acid ◽

Fermented Food ◽

Food Preservation ◽

Plant Materials ◽

Brown Sugar ◽

Major Nutrients ◽

Lifestyle Diseases

A plant-based paste fermented by lactic acid bacteria and yeast (fermented paste) was made from various plant materials. The paste was made of fermented food by applying traditional food-preservation techniques, that is, fermentation and sugaring. The fermented paste contained major nutrients (carbohydrates, proteins, and lipids), 18 kinds of amino acids, and vitamins (vitamin A, B1 B2, B6, B12, E, K, niacin, biotin, pantothenic acid, and folic acid). It contained five kinds of organic acids, and a large amount of dietary fiber and plant phytochemicals. Sucrose from brown sugar, used as a material, was completely resolved into glucose and fructose. Some physiological functions of the fermented paste were examined in vitro. It was demonstrated that the paste possessed antioxidant, antihypertensive, antibacterial, anti-inflammatory, anti-allergy and anti-tyrosinase activities in vitro. It was thought that the fermented paste would be a helpful functional food with various nutrients to help prevent lifestyle diseases.

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Identification of Listeria monocytogenes Genes Involved in Salt and Alkaline-pH Tolerance

Applied and Environmental Microbiology ◽

10.1128/aem.69.6.3137-3143.2003 ◽

2003 ◽

Vol 69 (6) ◽

pp. 3137-3143 ◽

Cited By ~ 24

Author(s):

Rozenn Gardan ◽

Pascale Cossart ◽

Jean Labadie

Keyword(s):

Salt Stress ◽

Listeria Monocytogenes ◽

Food Processing ◽

Food Preservation ◽

Alkaline Ph ◽

Sequence Comparisons ◽

Insertional Mutants ◽

Ph Tolerance ◽

Alkaline Tolerance ◽

Intergenic Regions

ABSTRACT The capacity of Listeria monocytogenes to tolerate salt and alkaline stresses is of particular importance, as this pathogen is often exposed to such environments during food processing and food preservation. We screened a library of Tn917-lacZ insertional mutants in order to identify genes involved in salt and/or alkaline tolerance. We isolated six mutants sensitive to salt stress and 12 mutants sensitive to salt and alkaline stresses. The position of the insertion of the transposon was located in 15 of these mutants. In six mutants the transposon was inserted in intergenic regions, and in nine mutants it was inserted in genes. Most of the genes have unknown functions, but sequence comparisons indicated that they encode putative transporters.

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Characterisation of antagonistic Bacillus paralicheniformis (strain EAL) by LC–MS, antimicrobial peptide genes, and ISR determinants

Antonie van Leeuwenhoek ◽

10.1007/s10482-020-01423-4 ◽

2020 ◽

Vol 113 (8) ◽

pp. 1167-1177

Author(s):

Hardik Naik Jinal ◽

Krishnan Sakthivel ◽

Natarajan Amaresan

Keyword(s):

Antimicrobial Peptide ◽

Bacillus Paralicheniformis

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Production of Novel Polygalacturonase from Bacillus paralicheniformis CBS32 and Application to Depolymerization of Ramie Fiber

Polymers ◽

10.3390/polym11091525 ◽

2019 ◽

Vol 11 (9) ◽

pp. 1525 ◽

Cited By ~ 3

Author(s):

Md. Saifur Rahman ◽

Yoon Seok Choi ◽

Young Kyun Kim ◽

Chulhwan Park ◽

Jin Cheol Yoo

Keyword(s):

Industrial Applications ◽

Fermented Food ◽

Ramie Fiber ◽

16S Rrna Sequence ◽

Application Study ◽

Glycosidic Bonds ◽

Bacillus Paralicheniformis ◽

Sds Page ◽

16S Rrna Sequence Analysis ◽

Terminal Amino

Polygalacturonase (EC. 3.2.1.15) is an enzyme that hydrolyzes the alpha-1,4 glycosidic bonds between galacturonic acid. In this study, an alkaline polygalacturonase producer Bacillus paralicheniformis CBS32 was isolated from kimchi (conventional Korean fermented food). The 16S rRNA sequence analysis of the isolated strain revealed that it was 99.92% identical to B. paralicheniformis KJ 16LBMN01000156. The polygalacturonase from B. paralicheniformis CBS32 was named PN32, and the purified PN32 showed a 16.8% yield and a 33-fold purity compared to the crude broth. The molecular mass, 110 kDa, was determined by SDS-PAGE, and the active band was confirmed by zymography analysis. The N-terminal amino acid sequence residues of PN32 were determined to be Gly–Val–Lys–Glu–Val–X–Gln–Thr–Phe. In the sequence comparison, PN32 was suggested as a novel polygalacturonase, since the sequence was not matched with the previous reports. In an application study, enzymatic depolymerization of ramie was performed for fiber degumming, and the result showed that the PN32 had a 28% higher depolymerization compared to the commercial pectinase. Overall, based on the results, PN32 has high potential for industrial applications.

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Stability and Activity of the Antimicrobial Peptide Leg1 in Solution and on Meat and Its Optimized Generation from Chickpea Storage Protein

Foods ◽

10.3390/foods10061192 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1192

Author(s):

Marie-Louise Heymich ◽

Showmika Srirangan ◽

Monika Pischetsrieder

Keyword(s):

Antimicrobial Peptide ◽

Incubation Temperature ◽

Standard Procedure ◽

Storage Conditions ◽

Food Preservation ◽

Food Storage ◽

Potassium Sorbate ◽

Food Grade ◽

E Coli ◽

Spoilage Bacteria

The antimicrobial peptide Leg1 (RIKTVTSFDLPALRFLKL) from chickpea legumin is active against spoilage bacteria, yeast, and mold. The present study tested its effectiveness under food storage conditions and examined options to obtain a food-grade agent. The minimum inhibitory concentration (MIC) of Leg1 against E. coli (62.5 µM) proved stable over seven days at 20 °C or 4 °C. It was not influenced by reduced pH (5.0 vs. 6.8), which is relevant in food such as meat. An incubation temperature of 20 °C vs. 37 °C reduced the MIC to 15.6/7.8 µM against E. coli/B. subtilis. With a minimum bactericidal concentration in meat of 125/15.6 µM against E. coli/B. subtilis, Leg1 is equivalently effective as nisin and 5000–82,000 times more active than sodium benzoate, potassium sorbate, or sodium nitrite. Replacing the counter-ion trifluoroacetate derived from peptide synthesis by the more natural alternatives acetate or chloride did not impair the activity of Leg1. As an alternative to chemical synthesis, an optimized protocol for chymotryptic hydrolysis was developed, increasing the yield from chickpea legumin by a factor of 30 compared to the standard procedure. The present results indicate that food-grade Leg1 could possibly be applicable for food preservation.

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Isolation and identification of lactic acid bacteria from vegetable-growing soils in Da Lat, Lam Dong

The Journal of Agriculture and Development ◽

10.52997/jad.1.04.2020 ◽

2020 ◽

Vol 19 (04) ◽

pp. 1-9

Author(s):

Thanh T. L. Bien

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Salmonella Typhi ◽

Fermented Food ◽

Isolation And Identification ◽

E Coli ◽

Level 3 ◽

Promote Plant Growth ◽

Control Disease ◽

Entire Margin

Lactic acid bacteria (LAB) have been used for decades in agriculture to improve soils, control disease and promote plant growth. LAB have been isolated from fermented food, milks and plants, however, a few studies of LAB from soils have been reported. This study aimed to isolate, screen and identify LAB from vegetable-growing soils collected from Da Lat (Lam Dong province). From 33 soil samples, 25 LAB isolates were selected on MRS agar supplemented with 1% CaCO3. The LAB isolates formed small, creamy white, convex, entire margin colonies, and were Gram-positive, catalase-negative and rod-shaped bacteria. Based on the acid-producing capacity, five LAB isolates (DT2, CT3, CC2, XL7 and S2) that produced clear zones around colonies due to the solubilization of CaCO3 with diameters ranged from 1.03 - 1.33 cm, and 11.8 - 14.3 mg/mL acid after 2-day incubation at 30oC. All selected LAB isolates showed the capacity to inhibit the growth of Fusarium oxysporum at level 1 (inhibitory rates in range of 10.66 - 19.96%), and Phytopthora sp. at level 3 (inhibitory rates in range of 50.86 - 57.44%) after 3 days. The isolates did not inhibit against E. coli and Staphylococcus but inhibit the growth of Bacillus spizizenii and Salmonella typhi with average inhibition diameters in range of 3.33 - 4.90 mm and 2.43 - 3.37 mm, respectively, after 1-day incubation. The five LAB isolates were molecularly determined to be Lactobacillus plantarum with 97 - 100% similarities

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Antimicrobial activities of Bacillus velezensis strains isolated from stingless bee products against methicillin-resistant Staphylococcus aureus

PLoS ONE ◽

10.1371/journal.pone.0251514 ◽

2021 ◽

Vol 16 (5) ◽

pp. e0251514

Author(s):

Mohamad Malik Al-adil Baharudin ◽

Mohamad Syazwan Ngalimat ◽

Fairolniza Mohd Shariff ◽

Zetty Norhana Balia Yusof ◽

Murni Karim ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Peptide ◽

High Performance ◽

Pathogenic Bacteria ◽

Antimicrobial Activities ◽

Proteinase K ◽

Tween 20 ◽

Bacillus Velezensis ◽

Promising Alternative ◽

Methicillin Resistant

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 μl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40–80°C), pH (4–12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.

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COCCUS BACTERIA IN FERMENTED DURIAN SPECIFIC FOOD OF BENGKULU

Jurnal Agroindustri ◽

10.31186/j.agroind.7.1.37-43 ◽

2017 ◽

Vol 7 (1) ◽

pp. 37-43

Author(s):

Hasanuddin Hasanuddin

Keyword(s):

Lactic Acid ◽

Organic Acid ◽

Fermented Food ◽

Food Preservation ◽

Laboratory Analysis ◽

Spontaneous Fermentation ◽

Traditional Markets ◽

Durio Zibethinus ◽

Specific Food

Coccus bactria is the cell of bacteria formed ball or ellip in 0,5 – 1µm diameter. Fermented durian is the fermented food prepared from spontaneous fermentation of durian (Durio zibethinus)with or without salt by wild bacteria. The sources of data in this study was laboratory analysis. Productions center of fermented durian, traditional markets were sampled for the experiment. Sampels were collected weekly in a month. Samples were analyzed microbiologically to determine and identify bacteria infermented durian. There were two species of coccus bacteria involved in fermented durian namely Pediococcus acidilacticiand Leuconostoc mesentroides. The specieseswere included in lactic acid bacteria.Lactic acid (C2H5COOH) is the organic acid that can serve as a food preservation

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Purification and Characterization of a Novel Cold Shock Protein-Like Bacteriocin Synthesized by Bacillus thuringiensis

Scientific Reports ◽

10.1038/srep35560 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 11

Author(s):

Tianpei Huang ◽

Xiaojuan Zhang ◽

Jieru Pan ◽

Xiaoyu Su ◽

Xin Jin ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Cold Shock ◽

Food Preservation ◽

Proteinase K ◽

Cold Shock Protein ◽

Purification And Characterization ◽

Large Conformational Change ◽

Proteinase K Treatment ◽

Ph Range

Abstract Bacillus thuringiensis (Bt), one of the most successful biopesticides, may expand its potential by producing bacteriocins (thuricins). The aim of this study was to investigate the antimicrobial potential of a novel Bt bacteriocin, thuricin BtCspB, produced by Bt BRC-ZYR2. The results showed that this bacteriocin has a high similarity with cold-shock protein B (CspB). BtCspB lost its activity after proteinase K treatment; however it was active at 60 °C for 30 min and was stable in the pH range 5–7. The partial loss of activity after the treatments of lipase II and catalase were likely due to the change in BtCspB structure and the partial degradation of BtCspB, respectively. The loss of activity at high temperatures and the activity variation at different pHs were not due to degradation or large conformational change. BtCspB did not inhibit four probiotics. It was only active against B. cereus strains 0938 and ATCC 10987 with MIC values of 3.125 μg/mL and 0.781 μg/mL, and MBC values of 12.5 μg/mL and 6.25 μg/mL, respectively. Taken together, these results provide new insights into a novel cold shock protein-like bacteriocin, BtCspB, which displayed promise for its use in food preservation and treatment of B. cereus-associated diseases.

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