Role of Conserved Histidine Residues in D-Aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6

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概要

• 論文の詳細を見る

• D-Aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (Alcaligenes A-6) was strongly inactivated by diethylpyrocarbonate (DEPC). An H67N mutant was barely active, with a k_<cat>/K_m 6.3×10^4 times lower than that of the recombinant wild-type enzyme, while the H67I mutant lost detectable activity. The H67N mutant had almost constant K_m, but greatly decreased k_<cat>. These results suggested that His67 is essential to the catalytic event. Both H69N and H69I mutants were overproduced in the insoluble fraction. The k_<cat>/K_m of H250N mutant was reduced by a factor of 2.5×10^4-fold as compared with the wild-type enzyme. No significant difference between H251N mutant and wild-type enzymes in the K_m and k_<cat> was found. The Zn content of H250N mutant was nearly half of that of wild-type enzyme. These results suggest that the His250 residue might be essential to catalysis via Zn binding.

• 社団法人日本農芸化学会の論文
• 2000-01-23

著者

• 酒井 謙二

  九大院生資環

• 酒井 謙二

  九州大学大学院農学研究院

• Sakai Kenji

  Department of Plant Resources, Kyushu University

• HAYASHI Shinichi

  Department of Diagnostic Pathology, Graduate School of Medicine and Pharmaceutical Sciences

• WAKAYAMA Mamoru

  Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University

• Hayashi S

  Nagoya Univ. Nagoya Jpn

• MORIGUCHI Mitsuaki

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• YADA Harutaka

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• KANDA Shunichi

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• YATSUDA Yukinori

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• Wakayama Mamoru

  Dep. Of Biotechnology Fac. Of Life Sciences Ritsumeikan Univ.

• Wakayama Mamoru

  Department Of Bioscience And Biotechnology Faculty Of Science And Engineering Ritsumeikan University

• Wakayama Mamoru

  Institute. For Chemical Research Kyoto University

• Moriguchi Mitsuaki

  Department Of Food And Bioscience Beppu University

• Moriguchi Mitsuaki

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Hayashi S

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Physics Nagoya University

• Yada Harutaka

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Mori Masatsugu

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Kanda Shunichi

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Environmental Chemistry And Engineering Faculty Of Engineering Oita University

• SAKAI Kenji

  Research Institute for Theoretical Physics Hiroshima University

• Yatsuda Yukinori

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Kanda Shun-ichi

  Department of Applied Chemistry, Faculty of Engineering, Oita University

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