An inducible 1-butanol dehydrogenase, a quinohaemoprotein, is involved in the oxidation of butane by 'Pseudomonas butanovora'

Citation
As. Vangnai et Dj. Arp, An inducible 1-butanol dehydrogenase, a quinohaemoprotein, is involved in the oxidation of butane by 'Pseudomonas butanovora', MICROBIO-UK, 147, 2001, pp. 745-756
Citations number
48
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Microbiology
Journal title
MICROBIOLOGY-UK
ISSN journal
1350-0872 → ACNP
Volume
147
Year of publication
2001
Part
3
Pages
745 - 756
Database
ISI
SICI code
1350-0872(200103)147:<745:AI1DAQ>2.0.ZU;2-I
Abstract
Butane-grown 'Pseudomonas butanovora' expressed two soluble alcohol dehydro genases (ADHs), an NAD(+)-dependent secondary ADH and an NAD(+) independent primary ADH. Two additional NAD+-dependent secondary ADHs could be detecte d when cells were grown on 2-butanol and lactate. The inducible NAD(+)-inde pendent 1-butanol dehydrogenase (BDH) of butane-grown cells was primarily r esponsible for 1-butanol oxidation in the butane metabolism pathway. BDH wa s purified to near homogeneity and identified as a quinohaemoprotein, conta ining, per mol enzyme, 1.0 mol pyrroloquinoline quinone (PQQ) and 0.25 mol haem c as prosthetic groups. BDH was synthesized as a monomer of approximat ely 66 kDa. It has a broad substrate range, including primary alcohols, sec ondary alcohols, aldehydes, C-4 diols and aromatic alcohols. It exhibited t he lowest K-m (7+/-1 muM) and highest k(cat)/K-m (72 x 10(4) M-1 s(-1)) val ue towards l-butanol. BDH exhibited ferricyanide-dependent ADH activity. Ca lcium ions (up to 10 mM) increased BDH activity substantially. Two BDH inte rnal amino acid sequences showed 73 and 62% identity and 83 and 66% similar ity, respectively, when compared with an amino acid sequence of ethanol deh ydrogenase from Comamonas testosteroni. The presence of the inducible BDH a nd secondary ADH may indicate that the terminal and subterminal oxidation p athways are involved in butane degradation of butane-grown 'P. butanovora'.