Cloning and Characterization of the Gene Encoding Pyrroloquinoline Quinone-dependent Poly(vinyl alcohol) Dehydrogenase of Pseudomonas sp. Strain VM15C
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概要
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A gene library of poly(vinyl alcohol) (PVA)-degrading Pseudomonas sp. strain VM15C was constructed in Escherichia coli with the vector pUC18. Screening of this library with a Chromogenic PVA dehydrogenase assay resulted in the isolation of a clone that carries the gene (pdh) for the PVA dehydrogenase, and the entire nucleotide sequence of its structural gene was determined. The gene encodes a protein of 639 amino acid residues (68,045 Da) and in the deduced amino acid sequence, some putative functional sites, a signal sequence, a heme c-binding site, and a PQQ-binding site, were detected. The amino acid sequence showed low similarity to other types of quinoprotein dehydrogenases. PVA dehydrogenase expressed in E. coli clones required PQQ. Ca^<2+>, and Mg^<2+> stimulated the activity. PVA-dependent heme c reduction occurred with exogenous PQQ in cell extracts of the E. coli clone. The PVA dehydrogenase in the E. coli clone was localized in the cytoplasm.
- 社団法人日本農芸化学会の論文
- 1996-07-23
著者
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Nishi K
Department Of Bioscience And Biotechnology Graduate School Of Bioresource And Bioenvironmental Scien
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SHIMAO MASAYUKI
Departments of Biotechnology Faculty of Engineering, Tottori University
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HARAYAMA Shigeaki
Marine Biotechnology Institute
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Tamogami T
Department Of Biotechnology Tottori University
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TAMOGAMI Tsuyoshi
Department of Biotechnology, Tottori University
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NISHI Koichi
Department of Biotechnology, Tottori University
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Nishi K
Department Of Biotechnology Tottori University
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Shimao Masayuki
Department Of Biotechnology Tottori University
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