1887

Abstract

was investigated with respect to polyhydroxybutyrate (PHB) biosynthesis. Polyhydroxyalkanoate (PHA) accumulation contributing to approximately 18% of the cell dry weight was obtained in the presence of glucose. Gas chromatography–mass spectrometry and gel permeation chromatography of the purified PHA showed that this polyester was solely composed of 3-hydroxybutyrate and had a weight average molar mass of 55×10 g mol and a polydispersity of 16. An ORF encoding a conserved, hypothetical protein which shared approximately 47% identity with the PHB synthase from was identified within the complete genomic sequence. This putative PHB synthase gene, , consisted of a 2019 nt stretch of DNA (encoding 673 aa residues), which encoded a PHB synthase with a molecular mass of approximately 73 kDa. This is currently the largest PHA synthase identified. The coding region was subcloned into vector pBBR1-JO2 under promoter control. The resulting plasmid, pQQ4, mediated PHB accumulation in the mutant PHB4 and recombinant JM109(pBHR69), which produced the β-ketothiolase and acetoacetyl-CoA reductase from , contributing to approximately 62% and 6% of cell dry weight, respectively. Functional expression of the coding region of was confirmed by immunoblotting and PHB synthase activity.

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2001-12-01
2020-01-17
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