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Volume 147, Issue 12

Polyhydroxybutyrate biosynthesis in : molecular characterization of the polyhydroxybutyrate synthase Free

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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Keyword(s): CDW, cell dry weight , GC–MS, gas chromatography–mass spectrometry , GPC, gel permeation chromatography , PHA depolymerase , PHA synthase , PHA, polyhydroxyalkanoate , PHB, polyhydroxybutyrate and polyhydroxyalkanoate
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2001-12-01
2024-04-25
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Polyhydroxybutyrate biosynthesis in Caulobacter crescentus: molecular characterization of the polyhydroxybutyrate synthase
Microbiology 147, 3353 (2001); https://doi.org/10.1099/00221287-147-12-3353
/content/journal/micro/10.1099/00221287-147-12-3353
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