Open AccessMolecular cloning of two ( R )‐specific enoyl‐CoA hydratase genes from Pseudomonas aeruginosa and their use for polyhydroxyalkanoate synthesis
Author(s) -
Tsuge Takeharu,
Fukui Toshiaki,
Matsusaki Hiromi,
Taguchi Seiichi,
Kobayashi Genta,
Ishizaki Ayaaki,
Doi Yoshiharu
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09013.x
Subject(s) - polyhydroxyalkanoates , escherichia coli , aeromonas caviae , biology , gene , biochemistry , plasmid , pseudomonadaceae , pseudomonas aeruginosa , pseudomonas , bacteria , chemistry , microbiology and biotechnology , genetics , vibrionaceae
Two Pseudomonas aeruginosa genes, termed phaJ1 Pa and phaJ2 Pa , homologous to the Aeromonas caviae ( R )‐specific enoyl‐CoA hydratase gene ( phaJ Ac ) were cloned using a PCR technique to investigate the monomer‐supplying ability for polyhydroxyalkanoate (PHA) synthesis from β‐oxidation cycle. Two expression plasmids for phaJ1 Pa and phaJ2 Pa were constructed and introduced into Escherichia coli DH5α strain. The recombinants harboring phaJ1 Pa or phaJ2 Pa 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 phaJ1 Pa and phaJ2 Pa products have the monomer‐supplying ability for PHA synthesis from β‐oxidation cycle.