Cloning and Sequencing of a Gene Encoding Nitrite Reductase from Paracoccus denitrificans and Expression of the Gene in Escherichia coli
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概要
- 論文の詳細を見る
A structural gene for nitrite reductase (nirS) was cloned from a denitrifying bacterium Paracoccus denitrificans into Escherichia coli MV1184. The coding sequence of nirS consisted of 1788 nucleotides and the value of the G+C content was 68%. This gene seemed to be in an operon structure. The size of nitrite reductase (NIR) was predicted to be 65.5 kDal from the amino acid sequence, which was similar to the value determined with purified NIR by SDS-polyacrylamide gel electrophoresis analysis. From hydrophathy analysis, the NIR from P. denitrificans seemed to be a periplamic enzyme. Crude extract from the recombinant E. coli barboring nirS and about 10 kbp of its downstream flanking region had significant activity of NO and N_2O formation from nitrite (NO_2^-), whereas crude extract from E. coli harboring only nirS had only weak activity. This result suggested that the downstream region includes the gene responsible for the protein which involved in NIR activation. In addition, the downstream region seemed to have a NO reductase gene, because NO to N_2O conversion activity was also detected in the crude extract from the recombinant E. coli harboring nirS and about 10 kbp of its downstream flanking region.
- 公益社団法人日本生物工学会の論文
- 1993-08-25
著者
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Uozumi N
Nagoya Univ. Nagoya Jpn
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Iijima Shinji
Department Of Biotechnology School Of Engineering Naogya University
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Iijima Shinji
Department Of Biotechnology Faculty Of Engineering Nagoya University
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Kobayashi Takeshi
Department Of Biological Chemistry College Of Bioscience And Biotechnology Chubu University
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Sugiyama Makoto
Department Of Radiology Hokuriku Hospital
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Uozumi Nobuyuki
Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University
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Ohshima T
Gunma Univ.
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Ohshima Takayuki
Department Of Biological And Chemical Engineering Gunma University
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Sugiyama Makoto
Department Of Biotechnology Faculty Of Engineering Nagoya University
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IIJIMA Shinji
Department of Agricultural Chemistry, The University of Tokyo
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