Expression and Characterization of ( R )-Specific Enoyl Coenzyme A Hydratase Involved in Polyhydroxyalkanoate Biosynthesis by Aeromonas caviae

• DOI: 10.1128/jb.180.3.667-673.1998
• Publication Date: 2019-12-31T16:30:16Z
• Authors: Toshiaki Fukui; Naofumi Shiomi; Yoshiharu Doi
• Keywords: Aeromonas caviae; Coenzyme A

ABSTRACT Complementation analysis of a polyhydroxyalkanoate (PHA)-negative mutant Aeromonas caviae proved that ORF3 in the pha locus (a 402-bp gene located downstream PHA synthase gene) participates biosynthesis on alkanoic acids, and is here referred to as phaJ Ac . Escherichia coli BL21(DE3) carrying under control T7 promoter overexpressed enoyl coenzyme A (enoyl-CoA) hydratase, which was purified by one-step anion-exchange chromatography. The N-terminal amino acid sequence hydratase corresponded deduced from nucleotide except for initial Met residue. enoyl-CoA encoded exhibited ( R )-specific hydration activity toward trans -2-enoyl-CoA with four six carbon atoms. These results have demonstrated 2-enoyl-CoA catalyzed translated product channeling pathway supplying )-3-hydroxyacyl-CoA monomer units fatty β-oxidation poly(3-hydroxybutyrate- co -3-hydroxyhexanoate) A. caviae.