Molecular Cloning and High Expression of the Bacillus Creatinase Gene in Escherichia coli

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概要

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• A genomic library of Bacillus sp. B-0618,prepared with the plasmid vector pACYC184,was screened to obtain a gene encoding creatinase (creatine amidinohydrolase; EC 3.5.3.3) by a birect coloration assay. A plasmid pCR1 isolated from a creatinase positive clone contained a 3.7-kb insert of the Bacillus chromosomal DNA. We determined the nucleotide sequence of a 1.55-kb segment containing the creatinase gene and found on open reading frame that coded for a protein consisting of 411 amino acids, with a calculated molecular mass of 46,946 daltons. The translated protein sequence of the creatinase gene from the Bacillus strain was 67% homologous to those of Pseudomonas and Flavobacterium. Although the creatinase of Bacillus sp. B-0618 was induced by choline chloride, Escherichia coli harboring pCR1 expressed 60-fold more creatinase activity intracellularly than did the producing strain, even in the absence of the inducer.

• 社団法人日本生物工学会の論文
• 1993-08-25

著者

• Sugiyama M

  Institute Of Pharmaceutical Sciences Hiroshima University School Of Medicine

• Sugiyama Masanori

  Institute Of Pharmaceutical Sciences Hiroshima University School Of Medicine

• SUZUKI KOJI

  Diagnostic Research and Developmemt Department, Diagnostic Division

• SAGAI HITOSHI

  Laboratory for Chemical Research, Institute for Life Science Research

• IMAMURA SHIGEYUKI

  Diagnostic Research and Development Department, Diafnostic Division

• IMAMURA Shigeyuki

  Central Laboratory, Asahi Chemical Industry Co., Ltd.

• Sugiyama Masanori

  Institute Of Pharmaceutical Sciences Faculty Of Medicine Hiroshima University

• Sagai Hitoshi

  Laboratory For Chemical Research Institute For Life Science Research:asahi Chemical Industry Co. Ltd

• Suzuki K

  Department Of Biotechnology Tottori University

• Imamura S

  Asahi Chemical Ind. Co. Ltd. Shizuoka

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