Purification and characterization of benzoate-CoA ligase from Magnetospirillum sp. strain TS-6 capable of aerobic and anaerobic degradation of aromatic compounds.
Benzoate-CoA ligase (EC 6.2.1.25), the initial enzyme of anaerobic benzoate degradation, was purified and characterized from Magnetospirillum sp. strain TS-6 grown under both anaerobic and aerobic conditions. The enzyme purified from anaerobically grown cells was a homodimer with a relative molecular mass of 120 kDa. The specific activity for benzoyl-CoA synthesis was 13.4 micromol min(-1) mg(-1) protein. The enzyme purified from aerobically grown cells was concluded to be the same gene product as the anaerobic enzyme. The benzoate-CoA ligase gene consisting of 1587 nucleotides was cloned and sequenced, and its induction under aerobic and anaerobic conditions during growth on benzoate was confirmed by quantitative reverse transcription PCR. These results indicate that a single benzoate-CoA ligase is expressed and benzoate is converted into benzoyl-CoA under both aerobic and anaerobic conditions in Magnetospirillum sp.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Kosuke
K
Kawaguchi
Yoshifumi
Y
Shinoda
Hiroya
H
Yurimoto
Yasuyoshi
Y
Sakai
Nobuo
N
Kato
D001565:Benzoates / Q000378:metabolism; D001673:Biodegradation, Environmental; D003066:Coenzyme A Ligases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D019281:Dimerization; D005798:Genes, Bacterial; D042201:Magnetospirillum / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction
D001565:Benzoates; D003066:Coenzyme A Ligases; C055455:benzoate coenzyme A ligase