Marine habitat harbors a wide variety of Bacillus species which are dominant producers of biologically active extracellular metabolites and anti-microbial peptides(AMPs).The present study reports a marine bacterial st...Marine habitat harbors a wide variety of Bacillus species which are dominant producers of biologically active extracellular metabolites and anti-microbial peptides(AMPs).The present study reports a marine bacterial strain MAH84 explored for production of a bioactive compound,its characterization and applications as antibiofilm agent.Cell free supernatant of strain MAH84(Nutrient broth amended with 1%Glucose),was purified by two resins[Diethylaminoethyl(DEAE)Sepharose and Sephadex G-25].The peptide mass fingerprinting(PMF)data of column purified cell free supernatant revealed it as zinc metalloproteases that belonged to M30 family of peptidases(hyicolysin)with a zinc motif at N-terminus with sequence AHEYQHM at position 102.Enzyme characterization studies showed that enzyme is thermostable with optimum activity at 60◦C,pH 12,halotolerance,and halo-stability(1-4 M NaCl).The enzyme activity in presence of chloride salts of metal[magnesium chloride(98.75%)and ferric chloride(97.5%)],and solvents[ethyl acetate(83.4%),glycol(98.4%),toluene(92%),methanol(99.6%)and acetone(93%)were found to be more than 80%.In continuation,application wise enzyme exhibited>50%antibiofilm activity against bacterial pathogens(such as E.coli MTCC43,P.aeruginosa MTCC424,K.pneumoniae MTCC9751,S.aureus MTCC96,and Salmonella enterica Subsp.Enterica serotype Abony-MTCC 6017),and high radical scavenging activity was noted i.e.,(2,2-diphenyl-1-picrylhydrazyl)free radical scavenging(DPPH Assay)82%,2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid(ABTS assay)69%,and Ferric reducing power activity(FRPA assay)69%.Enzyme was found to be non-cytotoxic towards HeLa cell lines at 10μg/ml.Additionally,a combinatorial study of ciprofloxacin,H2O2,short-chain fatty acids(acetic acid,propionic acid,and butyric acid)at their respective MIC(as determined)was conducted wherein the enzymes antibiofilm activity increased by 70%when co-administered with column purified cell free supernatant(metalloproteases 10μg/ml).Further,quantitative Reverse Transcriptase Polymerase Chain Reaction(q-RTPCR)revealed downregulation of two biofilm genes,namely csgD and bcsA in S.Abony with highest fold downregulation(2.48-fold)with butyric acid(100μg/ml)in concurrent administration with column purified cell free supernatant(10μg/ml).From our results,it is inferred that metalloprotease M30 from strain MAH84 can be a compelling quest for biofilm management.展开更多
Traditionally fermented fruit beverages have been consumed for centuries due to their pleasant sensory qualities,high nutritional value,and support for human health,particularly in resource-limited settings,compared t...Traditionally fermented fruit beverages have been consumed for centuries due to their pleasant sensory qualities,high nutritional value,and support for human health,particularly in resource-limited settings,compared to fresh fruit juices.While the therapeutic potential of traditional fermented beverages is increasingly recognized due to their diverse bioactive constituents,empirical evidence that supports these claims is still limited and requires further investigation.This study explores the nutritional and phytochemical characteristics of four variants of traditionally fermented pineapple-based beverages(TFBs):i.plain pineapple version(FPC),ii.amended with brown sugar(FPO),and two fruit-enriched types incorporating either iii.blueberry(FPB)or iv.pomegranate(FPP)alongside brown sugar after 72 h of fermentation.According to nutritional parameters,high content of Vitamin B12(4.07±0.01 mcg/100 mL)and Magnesium(6.73±0.02 mg/100 g)was observed in FPB relative to other fermented beverages.Moreover,FPB exhibited the highest phenolic(3.55±1.06 mg/mL),flavonoid content(3.77±0.28 mg/mL),and strongest antioxidant activity.Sugars were undetectable in FPC but present in FPO,FPB,and FPP,while fructose exhibited the most stable and predominant presence.GC-MS analysis revealed distinct variations in the composition and abundance of postbiotics across the four TFBs.Oleanolic acid exhibited the strongest interaction(-8.4 kcal/mol)against the COX-2 protein target 3LN1.This underscores 3LN1 as a preferable target for screening natural COX-2 inhibitors and supports the therapeutic relevance of traditionally fermented beverages through an integrated approach involving metabolomics and molecular docking.展开更多
基金financial support[EMR/2016/003586(SERB/F/942/2017–2020)]DST FIST,GoI Sanction order No.SR/FST/LSI-613/2014(C)Author PGSM thank UGC-GoI([202223-UGCES-22-GE-TEL-F-SJSGC(Savitribai Jyotirao Phule Single Girl Child)-1823]for fellowship.
文摘Marine habitat harbors a wide variety of Bacillus species which are dominant producers of biologically active extracellular metabolites and anti-microbial peptides(AMPs).The present study reports a marine bacterial strain MAH84 explored for production of a bioactive compound,its characterization and applications as antibiofilm agent.Cell free supernatant of strain MAH84(Nutrient broth amended with 1%Glucose),was purified by two resins[Diethylaminoethyl(DEAE)Sepharose and Sephadex G-25].The peptide mass fingerprinting(PMF)data of column purified cell free supernatant revealed it as zinc metalloproteases that belonged to M30 family of peptidases(hyicolysin)with a zinc motif at N-terminus with sequence AHEYQHM at position 102.Enzyme characterization studies showed that enzyme is thermostable with optimum activity at 60◦C,pH 12,halotolerance,and halo-stability(1-4 M NaCl).The enzyme activity in presence of chloride salts of metal[magnesium chloride(98.75%)and ferric chloride(97.5%)],and solvents[ethyl acetate(83.4%),glycol(98.4%),toluene(92%),methanol(99.6%)and acetone(93%)were found to be more than 80%.In continuation,application wise enzyme exhibited>50%antibiofilm activity against bacterial pathogens(such as E.coli MTCC43,P.aeruginosa MTCC424,K.pneumoniae MTCC9751,S.aureus MTCC96,and Salmonella enterica Subsp.Enterica serotype Abony-MTCC 6017),and high radical scavenging activity was noted i.e.,(2,2-diphenyl-1-picrylhydrazyl)free radical scavenging(DPPH Assay)82%,2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid(ABTS assay)69%,and Ferric reducing power activity(FRPA assay)69%.Enzyme was found to be non-cytotoxic towards HeLa cell lines at 10μg/ml.Additionally,a combinatorial study of ciprofloxacin,H2O2,short-chain fatty acids(acetic acid,propionic acid,and butyric acid)at their respective MIC(as determined)was conducted wherein the enzymes antibiofilm activity increased by 70%when co-administered with column purified cell free supernatant(metalloproteases 10μg/ml).Further,quantitative Reverse Transcriptase Polymerase Chain Reaction(q-RTPCR)revealed downregulation of two biofilm genes,namely csgD and bcsA in S.Abony with highest fold downregulation(2.48-fold)with butyric acid(100μg/ml)in concurrent administration with column purified cell free supernatant(10μg/ml).From our results,it is inferred that metalloprotease M30 from strain MAH84 can be a compelling quest for biofilm management.
基金supported by the University Grants Commission(UGC),Ministry of Education Government of India under the Savitribai Jyotirao Phule Single Girl Child Fellowship(SJSGC)to Prattipati Geya Sai Manjusha,Grant File no[F.No.82-7/2022(SA-III)]Registration ID:202223-UGCES-22-GE-TEL-F-SJSGC-1823Author’s also acknowledge the partial support from OU-DBT Builder,India(BT/INF/22/SP41415/2021,No:25/DBT-BUILDER/2023)。
文摘Traditionally fermented fruit beverages have been consumed for centuries due to their pleasant sensory qualities,high nutritional value,and support for human health,particularly in resource-limited settings,compared to fresh fruit juices.While the therapeutic potential of traditional fermented beverages is increasingly recognized due to their diverse bioactive constituents,empirical evidence that supports these claims is still limited and requires further investigation.This study explores the nutritional and phytochemical characteristics of four variants of traditionally fermented pineapple-based beverages(TFBs):i.plain pineapple version(FPC),ii.amended with brown sugar(FPO),and two fruit-enriched types incorporating either iii.blueberry(FPB)or iv.pomegranate(FPP)alongside brown sugar after 72 h of fermentation.According to nutritional parameters,high content of Vitamin B12(4.07±0.01 mcg/100 mL)and Magnesium(6.73±0.02 mg/100 g)was observed in FPB relative to other fermented beverages.Moreover,FPB exhibited the highest phenolic(3.55±1.06 mg/mL),flavonoid content(3.77±0.28 mg/mL),and strongest antioxidant activity.Sugars were undetectable in FPC but present in FPO,FPB,and FPP,while fructose exhibited the most stable and predominant presence.GC-MS analysis revealed distinct variations in the composition and abundance of postbiotics across the four TFBs.Oleanolic acid exhibited the strongest interaction(-8.4 kcal/mol)against the COX-2 protein target 3LN1.This underscores 3LN1 as a preferable target for screening natural COX-2 inhibitors and supports the therapeutic relevance of traditionally fermented beverages through an integrated approach involving metabolomics and molecular docking.
