Cloning of Cold-active Alkaline Phosphatase Gene of a Psychrophile, Shewanella sp., and Expression of the Recombinant Enzyme(Biochemistry & Molecular Biology)
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概要
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A psychrophilic alkaline phosphatase (EC 3. 1. 3. 1) from Shewanella sp. is a cold-active enzyme that has high catalytic activity at low temperature [Ishida et al. (1998) Biosci. Biotechnol. Biochem., 62, 2246-2250]. Here, we identified the nucleotide sequence of a gene encoding the enzyme after cloning with the polymerase chain reaction (PCR) and inverted PCR techniques. The deduced amino acid sequence of the enzyme contained conserved amino acids found among mesophilic alkaline phosphatases and showed some structural characteristics including a high content of hydrophobic amino acid residues and the lack of single α-helix compared with the alkaline phosphatase of Escherichia coli, which were possibly efficient for catalytic reaction at low temperatures. The recombinant enzyme expressed in E. coli was purified to homogeneity with the molecular mass of 41 kDa. The recombinant enzyme had a specific activity of 1,500 units/mg and had high catalytic activity at low temperatures.
- 社団法人日本農芸化学会の論文
- 2002-04-23
著者
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YAMAGATA Hiroshi
Kobe Univ.
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TSURUTA Hiroki
Laboratory of Biological Chemistry, Department of Biofunctional Chemistry, Faculty of Agriculture, K
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AIZONO Yasuo
Laboratory of Biological Chemistry, Department of Biofunctional Chemistry, Faculty of Agriculture, K
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Yamagata Hiroshi
Laboratory Of Biological Chemistry Faculty Of Agriculture Kobe University
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Yamagata Hiroshi
Laboratory Of Biochemistry Faculty Of Agriculture Kobe University
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Yamagata Hiroshi
Department Of Agricultural Chemistry College Of Agriculture Kyoto Prefectural University
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Aizono Y
Laboratory Of Biological Chemistry Department Of Biofunctional Chemistry Faculty Of Agriculture Kobe
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Aizono Yasuo
Lab. Of Biological Chemistry Dep. Of Biofunctional Chemistry Graduate School Of Agricultural Sci. Ko
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Aizono Yasuo
Laboratory Of Biological Chemistry Department Of Biofunctinal Chemistry Faculty Of Agriculture Kobe
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Tsuruta Hiroki
Center For Cooperative Research And Development Kobe University
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Tsuruta Hiroki
Laboratory Of Biological Chemistry Department Of Biofunctional Chemistry Faculty Of Agriculture Kobe
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MURAKAWA Takeshi
Laboratory of Biological Chemistry, Department of Biofunctional Chemistry, Faculty of Agriculture, K
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Murakawa Takeshi
Laboratory Of Biological Chemistry Department Of Biofunctional Chemistry Faculty Of Agriculture Kobe
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Tsuruta Hiroki
Laboratory of Biochemistry, Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University
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Aizono Yasuo
Laboratory of Biochemistry, Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University
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