Biosynthesis of medium-chain-length poly(hydroxyalkanoates) with altered composition by mutant hybrid PHA synthases |
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Authors: | Email author" target="_blank">Daniel?K?Y?SolaimanEmail author |
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Institution: | (1) Eastern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, PA 19038, USA |
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Abstract: | Pseudomonas resinovorans harbors two isogenic poly(hydroxyalkanoates) (PHAs) synthase genes (phaC1
Pre
, phaC2
Pre
) responsible for the production of intracellular medium-chain-length (mcl-)PHAs. Sequence analysis showed that the putative
gene-products of these genes contain a conserved α/β-hydrolase fold in the carboxy-terminal half of the proteins. Hybrid genes pha7 and pha8 were constructed by exchanging the α/β-hydrolase-fold coding portions of phaC1
Pre
and phaC2
Pre
at the 3′ terminal. When grown with decanoate as carbon source, the pha7- or pha8-transformed Escherichia coli LS1298 produced PHAs containing 73–75% β-hydroxydecanoate (β-HD) and 25–27% β-hydroxyoctanoate (β-HO). Deletion mutants,
Δpha7 and Δpha8, were isolated during the PCR-based construction of pha7 and pha8, respectively. Cells harboring these mutants produced PHAs containing 55–60 mol% β-HD and 40–45 mol% β-HO. These results
demonstrate the feasibility of generating active hybrid mcl-PHA synthase genes and their mutants with the potential of producing
polymers having a varied repeat-unit composition. |
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Keywords: | α /β -Hydrolase fold Hybrid gene Poly(hydroxyalkanoate) synthases |
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