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Title: Determination of genetic basis for biosurfactant production in distillery and curd whey wastes utilizing Pseudomonas aeruginosa strain BS2
Authors: Dubey, Kirti
Juwarkar, Asha
Keywords: biosurfactant
curd whey
curing
distillery waste
plasmid
Pseudomonas aeruginosa strain BS2
Issue Date: Jan-2004
Publisher: CSIR
Abstract: Pseudomonas aeruginosa strain BS2 has been demonstrated to have an ability to produce potent biosurfactant, an eco-friendly substitute to synthetic surfactants from distillery and whey wastes and capable of reducing the pollution load of these wastes in the range of 85-90%. To determine the basis for future identification of the genes responsible for biosurfactant production from wastes, studies on the presence of plasmid if any, its profile and role in biosurfactant production were performed. Suitable plasmid screening technique was selected because strain BS2 produced excessive slime in Luria Burnetti broth, which interfered with the migration and detection of plasmid. Among the several methods, alkaline lysis method was the most suitable which aided in recovery of slime-free cell lysate and resulted in the formation of a discrete band of plasmid in agarose gel. Plasmid profile study demonstrated that plasmid had high molecular weight of 32.08x106 Da and possessed the genetic determinants for antibiotics (chloramphenicol, tetracycline and sulphonamide) and heavy metal salt (mercuric chloride) resistance and were used as markers in curing experiment. To determine the role of megaplasmid in biosurfactant production, curing of megaplasmid was performed at highest sublethal doses of acridine orange (100 g/ml) and mitomycin-C (15 g/ml). Results indicated that only mitomycin-C treatment resulted in 28% of cell population which turned sensitive towards marker antibiotics and heavy metal salt due to loss of megaplasmid, which was further confirmed by agarose gel electrophoresis. Comparative analysis of biosurfactant production potential of cured cells with that of wild cells in both the wastes showed that the cured cells had similar potential capability of biosurfactant production as of wild strain which illustrates that genes responsible for biosurfactant production in distillery and whey wastes utilizing strain BS2 were not plasmid borne but resided on the chromosome where they are more stable.
Page(s): 74-81
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Source:IJBT Vol.03(1) [January 2004]

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