Cloning, Sequencing, High Expression, and Crystallization of the Thermophile Thermus aquaticus Glycerol Kinase
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概要
- 論文の詳細を見る
Glycerol kinase (EC 2.7.1.30) is a key enzyme of glycerol uptake and metabolism in bacteria. Using PCR, we amplified and cloned a glycerol kinase gene, glpK, from Thermus aquaticus. The complete gene has 1488 base pairs, coding for a protein of 496 amino acids with a predicted molecular weight of 54,814. The amino acid sequence deduced from T. aquaticus glpK was found to have identities of 97 and 81%, respectively, with those of Thermus flavus and Bacillus subtilis glpK genes. After overproduction in Escherichia coli, the expressed enzyme was easily purified to homogeneity by DEAE-Toyopearl chromatography. The purified enzyme has been crystallized by the hanging drop vapor diffusion method at 22℃. Comparison of the amino acid sequence with that of the B. subtilis enzyme showed that Ser and Lys are replaced by Ala and Arg, as was seen in mesophile and thermophile enzymes (Biochemistry, 18 : 5698-5703,1979).
- 社団法人日本農芸化学会の論文
- 1998-12-23
著者
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Kabashima T
School Of Pharmaceutical Sciences Nagasaki University
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Kabashima Tsutomu
School Of Pharmaceutical Sciences Nagasaki University
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吉田 豊和
岐阜大・工・生命工
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Ito Kiyoshi
School of Pharmaceutical Sciences, Nagasaki University
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Ito K
School Of Pharmaceutical Sciences Nagasaki University
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YOSHIMOTO TADASHI
School of Pharmaceutical Sciences, Nagasaki University
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Yoshimoto Tadashi
School Of Pharmaceutical Sciences Nagasaki University
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Ito K
Sapporo Breweries Ltd. Gumma Jpn
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Ito Kiyoshi
School Of Pharmaceutical Sciences Nagasaki University
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HUANG Hua-Shan
School of Pharmaceutical Sciences, Nagasaki University
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HUANG HuaShan
School of Pharmaceutical Sciences, Nagasaki University
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YIN ChangHong
School of Pharmaceutical Sciences, Nagasaki University
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Huang Huashan
School Of Pharmaceutical Sciences Nagasaki University
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Yin Changhong
School Of Pharmaceutical Sciences Nagasaki University
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Ito K
Yamagata Univ. Yonezawa Jpn
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Ito Kiyishi
School of Pharmaceutical Sciences, Nagasaki University
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Yin Chang-Hong
School of Pharmaceutical Sciences, Nagasaki University
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Yoshimoto Tadashi
School of Pharamaceutical Sciences, Nagasaki University
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