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Takeharu Tsuge, Toshiaki Fukui, Hiromi Matsusaki, Seiichi Taguchi, Genta Kobayashi, Ayaaki Ishizaki, Yoshiharu Doi, Molecular cloning of two (R)-specific enoyl-CoA hydratase genes from Pseudomonas aeruginosa and their use for polyhydroxyalkanoate synthesis, FEMS Microbiology Letters, Volume 184, Issue 2, March 2000, Pages 193–198, https://doi.org/10.1111/j.1574-6968.2000.tb09013.x
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Abstract
Two Pseudomonas aeruginosa genes, termed phaJ1Pa and phaJ2Pa, homologous to the Aeromonas caviae (R)-specific enoyl-CoA hydratase gene (phaJAc) were cloned using a PCR technique to investigate the monomer-supplying ability for polyhydroxyalkanoate (PHA) synthesis from β-oxidation cycle. Two expression plasmids for phaJ1Pa and phaJ2Pa were constructed and introduced into Escherichia coli DH5α strain. The recombinants harboring phaJ1Pa or phaJ2Pa showed high (R)-specific enoyl-CoA hydratase activity with different substrate specificities, that is, specific for short chain-length enoyl-CoA or medium chain-length enoyl-CoA, respectively. In addition, co-expression of these two hydratase genes with PHA synthase gene in E. coli LS5218 resulted in the accumulation of PHA up to 14–29 wt% of cell dry weight from dodecanoate as a sole carbon source. It has been suggested that phaJ1Pa and phaJ2Pa products have the monomer-supplying ability for PHA synthesis from β-oxidation cycle.