Cloning of the Gene for Inorganic Pyrophosphatase from a Thermoacidophilic Archaeon, Sulfolobus sp. Strain 7,and Overproduction of the Enzyme by Coexpression of tRNA for Arginine Rare Codon

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概要

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• The gene encoding an extremely stable inorganic pyrophosphatase from Sulfolobus sp. strain 7,a thermoacidophilic archaeon, was cloned and sequenced. An open reading frame consisted of 516 base pairs coding for a protein of 172-amino acid residues. The deduced sequence was supported by partial amino acid sequence analyses. All the catalytically important residues were conserved. A unique 17-base-pair sequence motif was found to be repeated four times in frame in the gene, encoding a cluster of acidic amino acids essential for the function. Although the codon usage of the gene was quite different from that of Escherichia coli, the gene was effectively expressed in E. coli. Coexpression of tRNA^<Arg>, cognate for the rare codon AGA in E. coli, however, further improved the production of the enzyme, which occupied more than 85% of the soluble proteins obatined after removal of heat denatured E. coli proteins.

• 社団法人日本農芸化学会の論文
• 1998-12-23

著者

• Oshima Tairo

  Department of Molecular Biology, School of Life Science, Tokyo University of Pharmacy and Life Scien

• WAKAGI Takayoshi

  Department of Biotechnology, The University of Tokyo

• Wakagi Takayoshi

  Department Of Biotechnology The University Of Tokyo

• IMAMURA Hiromi

  Department of Biotechnology, University of Tokyo

• Imamura H

  Yamaguchi Univ. Yamaguchi Jpn

• Imamura Hiromi

  Department Of Biotechnology University Of Tokyo

• Oshima Tairo

  Department Of Life Science Faculty Of Biocscience And Bioctechnology Tokyo Institute Of Technology

• Oshima T

  Department Of Molecular Biology Tokyo University Of Pharmacy And Life Science

• MATSUZAWA Hiroshi

  Department of Biotechnology, The University of Tokyo

• Wakagi Takayoshi

  Department Of Biotechnology Graduate School Of Agricultural And Life Sciences The University Of Toky

• Matsuzawa Hiroshi

  Department Of Biotechnology And Biotechnology Research Center The University Of Tokyo

• Matsuzawa Hiroshi

  Department of Bioscience and Biotechnology, Aomori University

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