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Cell Journal، جلد ۱۳، شماره Supplement، صفحات ۰-۰
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عنوان انگلیسی P-27: Isolation of Antimicrobial Peptidesfrom Vicia Faba and Determination of its Electrophoretic Pattern
چکیده انگلیسی مقاله Objective: Antimicrobial peptides (AMPs) are important substances functioning as self defense against infection by various harmful pathogens. They are isolated from various sources among animals, plants, and bacteria. Most of these characterized peptides are less than 10 kDa, have overall net positive charge, hydrophobic and exhibit bacterial, fungicidal, virucidal and tumoricidal properties. The present study shows the electrophoretic pattern of peptides from Vicia Faba seed and reports our first attempt to study the antibacterial activity of the extract on solid media and in broth media. Materials and Methods: 100 g seed were ground and protein was extracted in extraction buffer(10 mM Na2HPO4, 15 mM NaH2PO4, 100mM KCl, 2 mM EDTA, 2mM Thiurea 1.5% PVPP). Following a 70% ammonium sulfate precipitation. The pellets were resuspended in 50mM ammonium acetate and used for gel filtration chromatography (Sephadex G 75). The crude extract electrophoresis was carried out on 12% SDS- PAGE gel. Antibacterial activity on E.Coli and B. Subtilis from hospital infection was tested by incubating 105 cells in 70 μl of PB with 30 μl of peptide solutions or 30μl water as a control. After 2 hours at 25 ºC four serial dilutions were made, then 100 μl aliquots were plated on Mueller Hilnton plates. Colonies were counted. Also for agar well diffusion method, 105 cells were poured on plate. After 2 hours, 100 μl of peptide solutions were poured in the well. After incubation, the antibacterial activity was evaluated by measuring the inhibition zone diameter observed. Results: The SDS-PAGE gel show the crude extract contains many proteins with different spectrum molecular weight of proteins and peptides. Our experiment didn’t show any antibacterial activity on E.Coli and B. Subtilis from hospital infection. Conclusion: Antibacterial activity of crude extract should be tested on some standard bacteria.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/1559
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