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1.
Probiotics Antimicrob Proteins ; 12(4): 1571-1581, 2020 12.
Article in English | MEDLINE | ID: mdl-32385579

ABSTRACT

Bioactive peptides derived from chia (Salvia hispanica) seed with antioxidant, antihypertensive, and anti-inflammatory activities have been well documented; however, few studies describe the antimicrobial properties of these peptides, which is of great interest not only in the prevention of food-borne diseases but also food spoilage. The aim of this study was to generate chia seed peptides using microwave-assisted hydrolysis with sequential (alcalase + flavourzyme) enzymes (AF-MW), fractionate them into 3-10 and < 3 kDa fractions, and evaluate their potential antimicrobial activity towards Escherichia coli, Salmonella enterica, and Listeria monocytogenes. Overall, the peptide fraction < 3 kDa showed higher antimicrobial activity than both chia seed hydrolysate and peptide fraction 3-10 kDa. Furthermore, the < 3 kDa fraction showed remarkable increase in membrane permeability of E. coli (71.49% crystal violet uptake) and L. monocytogenes (80.10% crystal violet uptake). These peptides caused a significant extension in the lag phase, decreases in the maximum growth, and growth rate in the bacteria and promoted multiple indentations (transmembrane tunnels), membrane wrinkling, and pronounced deformations in the integrity of the bacterial cell membranes. Finally, a select group of peptides in the AF-MW < 3 kDa fraction contained 16 sequences with cationic and hydrophobic character, with seven of them sharing the exact same sequence (GDVIAIR) and eight of them having the amino acid K as either N- or C-terminal or both. In conclusion, our results indicate that bioactive peptides obtained from chia seed proteins by microwave and enzymatic hydrolysis could be employed as antimicrobial agents in foods and therapeutic applications.


Subject(s)
Anti-Bacterial Agents/pharmacology , Cell Membrane/drug effects , Plant Extracts/pharmacology , Plant Proteins/pharmacology , Protein Hydrolysates/pharmacology , Salvia/chemistry , Amino Acid Sequence , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Cell Membrane/ultrastructure , Cell Membrane Permeability/drug effects , Chemical Fractionation/methods , Endopeptidases/chemistry , Escherichia coli/drug effects , Escherichia coli/growth & development , Escherichia coli/ultrastructure , Hydrolysis , Listeria monocytogenes/drug effects , Listeria monocytogenes/growth & development , Listeria monocytogenes/ultrastructure , Microwaves , Plant Extracts/chemistry , Plant Proteins/chemistry , Plant Proteins/isolation & purification , Protein Hydrolysates/chemistry , Salmonella enterica/drug effects , Salmonella enterica/growth & development , Salmonella enterica/ultrastructure , Seeds/chemistry , Subtilisins/chemistry
2.
J Microbiol ; 52(8): 667-74, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24994011

ABSTRACT

In the present study, silver nanoparticles (AgNPs) with an average particle size of 5.5 ± 3.1 nm were biosynthesized using an endophytic fungus Cryptosporiopsis ericae PS4 isolated from the ethno-medicinal plant Potentilla fulgens L. The nanoparticles were characterized using UV-visible spectrophotometer, transmission electron microscopy (TEM), scanning electron microscopy (SEM), selective area electron diffraction (SAED), and energy dispersive X-ray (EDX) spectroscopy analysis. Antimicrobial efficacy of the AgNPs was analyzed singly and in combination with the antibiotic/antifungal agent chloramphenicol/fluconazole, against five pathogenic microorganisms--Staphylococcus aureus MTCC96, Salmonella enteric MTCC735, Escherichia coli MTCC730, Enterococcus faecalis MTCC2729, and Candida albicans MTCC 183. The activity of AgNPs on the growth and morphology of the microorganisms was studied in solid and liquid growth media employing various susceptibility assays. These studies demonstrated that concentrations of AgNPs alone between 10 and 25 µM reduced the growth rates of the tested bacteria and fungus and revealed bactericidal/fungicidal activity of the AgNPs by delaying the exponential and stationary phases. Examination using SEM showed pits and ruptures in bacterial cells indicating fragmented cell membrane and severe cell damage in those cultures treated with AgNPs. These experimental findings suggest that the biosynthesized AgNPs may be a potential antimicrobial agent.


Subject(s)
Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Antifungal Agents/isolation & purification , Antifungal Agents/pharmacology , Fungi/metabolism , Metal Nanoparticles , Silver , Candida albicans/drug effects , Candida albicans/growth & development , Candida albicans/ultrastructure , Chloramphenicol/pharmacology , Enterococcus faecalis/drug effects , Enterococcus faecalis/growth & development , Enterococcus faecalis/ultrastructure , Escherichia coli/drug effects , Escherichia coli/growth & development , Escherichia coli/ultrastructure , Fluconazole/pharmacology , Fungi/isolation & purification , Metal Nanoparticles/chemistry , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Salmonella enterica/drug effects , Salmonella enterica/growth & development , Salmonella enterica/ultrastructure , Spectrometry, X-Ray Emission , Spectrophotometry , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development , Staphylococcus aureus/ultrastructure
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