Isolation and Characterization of an Alcohol Dehydrogenase Gene from the Octylphenol Polyethoxylate Degrader Pseudomonas putida S-5
Octylphenol polyethoxylate (OPEO<SUB>n</SUB>) biodegradation by <I>Pseudomonas putida</I> S-5 under aerobic conditions is initiated by the oxidation of its terminal alcohol group by alcohol dehydrogenase. A DNA fragment, containing an alcohol dehydrogenase gene (<I>adh1</I>), was isolated using a combination of degenerate PCR and inverse PCR. The predicted translation product of <I>adh1</I> showed significant sequence similarity to bacterial alcohol dehydrogenases. Furthermore, a flavin-binding motif and signature patterns conserved in type III FAD-dependent alcohol oxidases were detected. Two open reading frames (ORFs) were found upstream of <I>adh1</I>, encoding a putative acyl-CoA synthetase and a putative esterase. Downstream of <I>adh1</I> and located on the opposite strand was an ORF encoding a putative aldehyde dehydrogenase. Transcription analysis using RT-PCR showed that <I>adh1</I> is cotranscribed with the putative acyl-CoA synthetase and esterase genes during growth on OPEO<SUB>n</SUB>. ADH1 overproduced in <I>Escherichia coli</I> exhibited activity not only toward various alcohols, including OPEO<SUB>n</SUB>s, but also toward primary aliphatic and aromatic aldehydes.
- Bioscience, biotechnology, and biochemistry
Bioscience, biotechnology, and biochemistry 70(8), 1855-1863, 2006-08-23
Japan Society for Bioscience, Biotechnology, and Agrochemistry