Synthesis of Short-chain-length/Medium-chain-length Polyhydroxyalkanoate (PHA) Copolymers in Peroxisome of the Transgenic Arabidopsis Thaliana Harboring the PHA Synthase Gene from Pseudomonas sp. 61-3
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In this paper, the photosynthetic production of short-chain-length/medium-chain-length polyhydroxyalkanoate (PHA) copolymers is reported. The wild-type and highly active doubly mutated PHA synthase 1 (S325T/Q481K, abbreviated ST/QK) genes from Pseudomonas sp. 61-3 were introduced into Arabidopsis thaliana. Peroxisome targeting signal 1 (PTS1) was used to target PHA synthases into the peroxisome to synthesize PHA from the intermediates of the β-oxidation pathway. The transgenic Arabidopsis produced PHA copolymers consisting of 40–57 mol% 3-hydroxybutyrate, 21–49 mol% 3-hydroxyvalerate, 8–18 mol% 3-hydroxyhexanoate, and 2–8 mol% 3-hydroxyoctanoate. The maximum PHA contents were 220μ g/g cell dry weight (cdw) in leaves, and 36μ g/g cdw in stems, respectively. The expression of the ST/QK mutated PHA synthase in leaves gene did not lead to significant difference in PHA content and monomer composition of PHAs, compared to the wild-type PHA synthase gene, suggesting that the supply of monomers may be a rate-determining step of PHA biosynthesis in the peroxisome. However, in stems, there were significant differences dependent on whether the wild-type or ST/QK mutated PHA synthase was expressed. These results suggest that tissue-specific monomer availability is important in determining the final mol% composition of PHA copolymers produced by the peroxisome in plants.
KeywordsPolyhydroxyalkanoate Arabidopsis thaliana Peroxisome Pseudomonas sp. 61-3 3-hydroxyvalerate
We thank Ms. Hiromi Masaki for supporting plant manipulations, Dr. Christopher Nomura for valuable discussions and RIKEN Research Resources Center (RRC) for DNA sequence analysis. This work was partly supported by Grant-in-aid for Scientific Research of Japan (Grant No. 16710059 to K.M.), Special Postdoctoral Research Program of RIKEN Institute (to K.M. and K.T.), Solution Oriented Research for Science, Technology (SORST) of the Japan Science and Technology Corporation (JST), Hokkaido Foundation for the Promotion of Scientific and Industrial Technology, and Industrial Technology Research Grant Program in 2003 from the New Energy and Industrial Technology Development Organization (NEDO).
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