Purification and Characterization of _L-Aminoacylase from Pseudomonas maltophila B1

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概要

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• The constitutive _L-aminoacylase, which is used for optical resolution of _<DL-α>-aminosuberic acid(_<DL>-Asu), has been purified and characterized from Pseudomonas maltophila B1. The crude enzyme showed a specific activity of 0.062 units/mg for N-acetyl(Ac)-_L-Asu. This value is very high compared with those from Aspergillus melleus, porcine kidney, and Bacillus stearothermophilus. Molecular masses of 108 kDa for the native enzyme and 50 kDa for the subunit were determined, indicating a dimer. The enzyme activity was optimal at pH 8.0 and at 55℃. The enzyme hydrolyzed N-acyl derivatives of various neutral _L-amino acids and acidic _L-amino acids, _L-glutamate and _L-Asu. The enzyme also had dipeptidase activity. The K_m values for N-Ac-_L-alanine and N-Ac-_<DL>-Asu were determined at 2.32 and 12.7mM, respectively. The apoenzyme was activated using Zn^<2+> Ca<2+> and Co<2+>. Glyoxylate, _<DL>-lactate, phenylboronic acid(PBA), butaneboronic acid(BBA), diethylpyrocarbonate(DEP), and phenylglyoxal(PGO)inhibited enzyme activity.

• 社団法人日本生物工学会の論文
• 1998-03-25

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

• WAKAYAMA Mamoru

  Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University

• SHIBA Eiichi

  Department of Surgical Oncology, Osaka University Medical School

• MORIGUCHI Mitsuaki

  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

• Shiba Eiichi

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Physics Nagoya University

• Mori Masatsugu

  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

• Sakai Kenji

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Shiiba Eiichi

  Department of Applied Chemistry, Faculty of Engineering, Oita University

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