Purification and Characterization of Thermostable Glycerol Kinase from Thermus flavus

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概要

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• Glycerol kinase (EC 2.7.1.30; ATP : glycerol 3-phosphotransferase) was purified from Thermus flavus, by ammonium sulfate fractionation and sequential chromatographies on Toyopearl HW65C and DEAE-Toyopearl columns, with an activity recovery of 22.7%. The enzyme is most active at pHs of 9.0 to 9.5. The optimm temperature fot the enzyme is 50-70℃. About 50% of the initial activity remains after incubation at 68℃ and pH 7.5 or 30 min. The isoelectric point of the enzyme is 4.3. Its molecular weight is estimted to be 220,000 Da by gel filtration on FPLC-Hiload Superdex 200 pg and 58,000 Da by SDS-PAGE, suggesting that it is a tetramer. The activity of the enzyme is completely inhibited by PCMB, HgCl_2 and Mn^<2+>. The K_m values of the enzyme for glycerol and ATP are 3.8×10^<-5>M and 1.62×10^<-4>M, resoectively. The N-terminal amino acid sequence of the enzyme is MNQYMLAIDQGTTSSR.

• 社団法人日本生物工学会の論文
• 1997-04-25

著者

• Kabashima T

  School Of Pharmaceutical Sciences Nagasaki University

• Kabashima Tsutomu

  School Of Pharmaceutical Sciences Nagasaki University

• 吉田 豊和

  岐阜大・工・生命工

• Ito Kiyoshi

  School of Pharmaceutical Sciences, Nagasaki University

• Ito K

  School Of Pharmaceutical Sciences Nagasaki University

• YOSHIMOTO TADASHI

  School of Pharmaceutical Sciences, Nagasaki University

• Yoshimoto Tadashi

  School Of Pharmaceutical Sciences Nagasaki University

• Ito K

  Sapporo Breweries Ltd. Gumma Jpn

• Meng Yang

  School Of Pharmaceutical Sciences Nagasaki University And Toyobo Tsuruga Institute Of Biotechnology

• Ito Kiyoshi

  School Of Pharmaceutical Sciences Nagasaki University

• HUANG Hua-Shan

  School of Pharmaceutical Sciences, Nagasaki University

• YOSHIDA TOMOHIRO

  School of Pharmaceutical Sciences, Nagasaki University, and Toyobo Tsuruga Institute of Biotechnolog

• NISHIYA YOSHIAKI

  School of Pharmaceutical Sciences, Nagasaki University, and Toyobo Tsuruga Institute of Biotechnolog

• KAWAMURA YOSHIHISA

  School of Pharmaceutical Sciences, Nagasaki University, and Toyobo Tsuruga Institute of Biotechnolog

• Huang Huashan

  School Of Pharmaceutical Sciences Nagasaki University

• Nishiya Yoshiaki

  School Of Pharmaceutical Sciences Nagasaki University And Toyobo Tsuruga Institute Of Biotechnology

• Kawamura Yoshihisa

  School Of Pharmaceutical Sciences Nagasaki University And Toyobo Tsuruga Institute Of Biotechnology

• Yoshida Tomohiro

  School Of Pharmaceutical Sciences Nagasaki University And Toyobo Tsuruga Institute Of Biotechnology

• Ito K

  Yamagata Univ. Yonezawa Jpn

• Yoshimoto Tadashi

  School of Pharamaceutical Sciences, Nagasaki University

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