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A lower specificity PhaC2 synthase from Pseudomonas stutzeri catalyses the production of copolyesters consisting of short-chain-length and medium-chain-length 3-hydroxyalkanoates

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Abstract

A polyhydroxyalkanoate (PHA) synthase gene phaC2 Ps from Pseudomonas stutzeri strain 1317 was introduced into a PHA synthase gene phbC Re negative mutant, Ralstonia eutropha PHB−4. It conferred on the host strain the ability to synthesize PHA, the monomer compositions of which varied widely when grown on different carbon sources. During cultivation on gluconate, the presence of phaC2 Ps in R. eutropha PHB−4 led to the accumulation of polyhydroxybutyrate (PHB) homopolymer in an amount of 40.9 wt% in dry cells. With fatty acids, the recombinant successfully produced PHA copolyesters containing both short-chain-length and medium-chain-length 3-hydroxyalkanoate (3HA) of 4–12 carbon atoms in length. When cultivated on a mixture of gluconate and fatty acid, the monomer composition of accumulated PHA was greatly affected and the monomer content was easily regulated by the addition of fatty acids in the cultivation medium. After the (R)-3-hydroxydecanol-ACP:CoA transacylase gene phaG Pp from Pseudomonas putida was introduced into phaC2 Ps-containing R. eutropha PHB−4, poly(3HB-co-3HA) copolyester with a very high 3-hydroxybutyrate (3HB) fraction (97.3 mol%) was produced from gluconate and the monomer compositions of PHA synthesized from fatty acids were also altered. This study clearly demonstrated that PhaC2Ps cloned from P. stutzeri 1317 has extraordinarily low substrate specificity in vivo, though it has only 54% identity in comparison to a previously described low-substrate-specificity PHA synthase PhaC1Ps from Pseudomonas sp. 61–3. This study also indicated that the monomer composition and content of the synthesized PHA can be effectively modulated by controlling the addition of carbon sources or by modifying metabolic pathways in the hosts.

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Acknowledgments

This research was supported by the key project No. 20334020 from the Natural Science Foundation of China (NSFC). The NSFC outstanding young scientist grant No. 30225001 awarded to CGQ also provided financial contribution for this study. We are grateful to Prof. A. Steinbüchel of the University of Münster in Germany for the generous donation of strain R. eutropha PHB−4, and to Dr. Phil Green of P&G Company, USA, for kindly providing us with plasmid pBBR1MCS2.

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  1. MOE Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China

    Jing-Yu Chen, Ge Song & Guo-Qiang Chen

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  1. Jing-Yu Chen
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  2. Ge Song
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Correspondence to Guo-Qiang Chen.

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Chen, JY., Song, G. & Chen, GQ. A lower specificity PhaC2 synthase from Pseudomonas stutzeri catalyses the production of copolyesters consisting of short-chain-length and medium-chain-length 3-hydroxyalkanoates. Antonie Van Leeuwenhoek 89, 157–167 (2006). https://doi.org/10.1007/s10482-005-9019-9

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  • DOI: https://doi.org/10.1007/s10482-005-9019-9

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