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Production of Short-Chain-Length/Medium-Chain-Length Polyhydroxyalkanoate (PHA) Copolymer in the Plastid of Arabidopsis thaliana Using an Engineered 3-Ketoacyl-acyl Carrier Protein Synthase III

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posted on 13.04.2009, 00:00 by Ken’ichiro Matsumoto, Takaaki Murata, Rina Nagao, Christopher T. Nomura, Satoshi Arai, Yuko Arai, Kazuma Takase, Hideo Nakashita, Seiichi Taguchi, Hiroaki Shimada
Short-chain-length/medium-chain-length (SCL/MCL) polyhydroxyalkanoate (PHA) was produced in the plastids of Arabidopsis thaliana. Phe87Thr (F87T) mutated 3-ketoacyl-acyl carrier protein (ACP) synthase III (FabH) from Escherichia coli, and Ser325Thr/Gln481Lys (ST/QK) mutated polyhydroxyalkanoate (PHA) synthase (PhaC1) from Pseudomonas sp. 61−3, along with the β-ketothiolase (PhaA) and acetoacetyl-CoA reductase (PhaB) from Ralstonia eutropha (Cupriavidus necator) genes were introduced into Arabidopsis. The transgenic Arabidopsis produced PHA copolymers composed of monomers consisting of 4−14 carbons. The introduction of the engineered PHA synthase resulted in a 10-fold increase in PHA content compared to plants expressing the wild-type PHA synthase. In addition, expression of the engineered fabH gene in the plastid led to an increase in the amount of the SCL monomer, 3-hydroxybutyrate, incorporated into PHA, and contributed to supply of MCL monomers for PHA production.

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