Cloning and Sequencing of a Gene Encoding D-Aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 and Expression of the Gene in Escherichia coli

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概要

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• The gene encoding the D-aminoacylase of Alcaligenes xylosoxydans subsp. xylosoxydans A-6(Alcaligenes A-6) was cloned and its complete nucleotide sequence was identified. The D-aminoacylase structural gene consists of 1452 nucleotides and encodes 484 amino acid residues. The molecular weight of D-aminoacylase was calculated to be 51,918. This value agreed well with the apparent molecular weight of 52,000 found for the purified enzyme from Alcaligenes A-6 by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE). The N-terminal amino acid sequence (NH_2-SQSDSQPFDLLRAG-) predicted by the nucleotide sequence exactly matched those of the purified D-aminoacylase both from Alcaligenes A-6 and from cloned Escherichia coli (E. coli), with the exception of the removal of the N-terminal methionine processed after translation. The purified recombinant enzyme showed almost the same enzymatic properties as the native enzyme from Alcaligenes A-6. Alcaligenes A-6 D-aminoacylase showed 25-29% homology with L-aminoacylases from Bacillus stearothermophilus, porcine, and humans.

• 社団法人日本農芸化学会の論文
• 1995-11-23

著者

• 酒井 謙二

  九大院生資環

• Sakai Kenji

  Department of Plant Resources, Kyushu University

• WAKAYAMA Mamoru

  Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University

• Miura Y

  Department Of Applied Biological Science Tokyo Noko University

• Miura Yutaka

  Applied Bioresearch Center Kirin Brewry Co. Ltd.

• Miura Yutaka

  Department Of Agricultural Chemistry Faculty Of Agriculture The University Of Tokyo

• Hayashi S

  Nagoya Univ. Nagoya Jpn

• HAYASHI Shin-Ichi

  Department of Immunology, School of Life Science Tottori University

• MORIGUCHI Mitsuaki

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• Miyamoto Yoshiro

  Department Of Biochemical Engineering Faculty Of Computer Science And System Engineering Kyushu Inst

• Wakayama Mamoru

  Department Of Bioscience And Biotechnology Faculty Of Science And Engineering Ritsumeikan 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

• Katsuno Yutaka

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• KATSUNO Yuko

  School of Food and Nutritional Sciences, University of Shizuoka

• Mori Masatsugu

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Environmental Chemistry And Engineering Faculty Of Engineering Oita University

• Hayashi Shin-ichi

  Department Of Immunology School Of Life Science Tottori University

• SAKAI Kenji

  Research Institute for Theoretical Physics Hiroshima University

• Miura Y

  Tokyo Noko Univ. Tokyo Jpn

• Miura Yutaka

  Dep. Of Applied Biological Sci. Tokyo Noko Univ.

• Sakai Kenji

  Department Of Drug Metabolism And Pharmacokinetics Nippon Boehringer Ingelheim Co. Ltd.

• Miyamoto Yoko

  Laboratory Of Biochemistry Faculty Of Agriculture Kyushu University

• Sakai Kenji

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Wakayama Mamoru

  Department of Applied Chemistry, Faculty of Engineering, Oita University

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