Monday, November 9, 2009
P42

Coproduction of biopolymers consisting of medium chain length 3-hydroxyalkanoic acid and exopolysaccharide by Pseudomonas CMG607w

Nazia Jamil1, Nuzhat Ahmed2, David H. Edwards3, Geoffrey M. Gadd4, and Shahida Hasnain1. (1) Microbiology & Molecular Genetics, University of the Punjab, New campus, Punjab University, Lahore, 54590, Pakistan, (2) Centre for Molecular Genetics, University of Karachi, Karachi-75270, Karachi, 75270, Pakistan, (3) Department of Molecular and Cellular Medicine, University of Dundee, Scotland, DD1 4HN, United Kingdom, (4) Division of Environmental and Applied Biology Biological Sciences Institute School of Life Sciences, University of Dundee Dunde, Scotland, DD1 4HN, United Kingdom

Metabolite production was analyzed in the supernatant of fermented broths of ASW (supplanted with different carbon sources), it was revealed that in presence of these nutrient conditions CMG607w possess metal binding activity, which may be due to the presence of a compound MW 349 and 41.11kDa protein in the supernatant, detected by GC/MS and phast gel analysis respectively, while EPS production was observed in presence of different carbon sources. Bioplastic (medium chain length polyhydroxylakanote) was extracted and purified from CMG607w. PHA synthesis was substrate depended in CMG607w. In presence of sodium gloconate spherical PHA granules were extruded from the cells, while in presence of sodium acetate, PHA granules were in form of crystals which make the cell fragile and releases from the cell by lyses of the cell membrane. Under highly enriched conditions, co production of polysaccharide and blends of PHB/PHA were observed. PCR based strategy was used to amplify Pha biosynthsis operon from chromosomal DNA. In CMG607w Pha biosynthsis operon has PhaCIZC2D (polymerasel, depolymerase, polymerase2 and hypothetical protein) genes orientation. Conserved sequences were observed in polymerase C1 and C2. All genes of Pha synthase operon were cloned and sequenced. Pha biosynthesis operon of CMG607w has PhaCIZC2D genes and have 98% homology to P. aeruginosa (AE004919, X66592), 87% to P. oleovorans (AF318050) and 77% Burkholderia caryphylli (AF394660) Genbank accession numbers for Pha biosynthesis locus present in CMG607w are ORF1(AY596791), PhaC1((AY5996788), PhaZ (AY596789) PhaC2 (AY5967790) and PhaD (AY596791).

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