The gene pvaB encodes oxidized polyvinyl alcohol hydrolase of Pseudomonas sp. strain VM15C and forms an operon with the polyvinyl alcohol dehydrogenase gene pvaA.
A 5.7 kbp SphI fragment containing the polyvinyl alcohol (PVA) dehydrogenase gene pvaA and its 1.9 kbp 5'-flanking region was cloned from the PVA-degrading bacterium Pseudomonas sp. VM15C. The pvaB gene, encoding oxidized PVA hydrolase, was found in the region upstream of pvaA. Sequence data and expression studies indicated that pvaA and B constitute an operon in the order pvaBA. The pvaB gene encoded a protein of 379 amino acid residues (40610 Da), and a lipoprotein signal sequence and the lipase consensus sequence, Gly-X-Ser-X-Gly, characteristic of the active-site serine region in serine hydrolases, were detected in the deduced amino acid sequence. The pvaB product with the pvaA product constituted an enzyme system for the cleavage of PVA molecules. The pvaA product introduced beta-diketone groups into the PVA molecule, and the pvaB product hydrolysed these beta-diketone groups in oxidized PVA. The pvaB product also hydrolysed 4,6-nonanedione at a low rate, but not acetylacetone or 5-nonanone. It was completely inhibited by PMSF and was concluded to be a serine hydrolase. There were no proteins showing high similarity to the pvaB product in the databases, but minor similarity to a number of serine hydrolases including polyhydroxyalkanoate depolymerases was apparent.
Shimao
Masayuki
M
Department of Biotechnology, Tottori University, Tottori, Tottori 680-8552, Japan1.
Tamogami
Tsuyoshi
T
Department of Biotechnology, Tottori University, Tottori, Tottori 680-8552, Japan1.
Kishida
Shinsuke
S
Department of Biotechnology, Tottori University, Tottori, Tottori 680-8552, Japan1.
Harayama
Shigeaki
S
Marine Biotechnology Institute, Kamaishi Laboratory, Kamaishi, Iwate 026-0001, Japan2.
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins; D001673:Biodegradation, Environmental; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics*; D010084:Oxidation-Reduction; D011142:Polyvinyl Alcohol / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
D001426:Bacterial Proteins; D011142:Polyvinyl Alcohol; D000429:Alcohol Oxidoreductases; C048312:polyvinyl-alcohol dehydrogenase (acceptor); D002265:Carboxylic Ester Hydrolases; C409492:oxidized polyvinyl alcohol hydrolase, Pseudomonas sp.