Efficient Removal of Sulfide Following Integration of Multiple Copies of the Sulfide-Quinone Oxidoreductase Gene(sqr)into the Escherichia coli Chromosome

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概要

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• For the oxidation and removal of hydrogen sulfide, which causes an offensive odor from the contents of animal intestines, recombinant strains of Escherichia coli were constructed. The sulfide-quinone oxidoreductase gene(sqr)from Rhodobacter capsulatus was integrated in low copy numbers into the chromosome of Escherichia coli W3110. Multiple copies of sqr on plasmids were also delivered into the cytoplasm of the same strain. The sqr genes were homologously transducted onto the chromosomal lacZ region and their existence there was verified by Southern blot analysis. Sulfide oxidation in a chemical medium effectively increased for the recombinant strains which carried 2〜3 copies of sqr under the control of the lac or tac promoter in the chromosome, and also for strains which carried 10 copies of sqr under the control of the lac or tac promoter on plasmids. In both types of recombinant, the tac promoter was more effective for SQR expression than the lac promoter. Construction of a recombinant with 3 copies of sqr under the control of the tac promoter in the chromosome was unsuccessful. In recombinants with SQR activity lower than 700 nmol/mg cell protein/min, oxygen consumption increased proportionally to SQR activity. An elevation in SQR activity in this range resulted in an increase in oxygen consumption and a decrease in sulfide concentration. When the recombinant cells were cultured until the 160th generation, WL2, WL3 and WT2, which carried 2, 3 and 2 copies of sqr in the chromosome, respectively, retained SQR activity similar to that of the first generation. For WL300 and WT20 which carried multi-copies of sqr in plasmids SQR activity was undetectable. The recombinant with 2 copies of sqr in the chromosome regulated by the tac promoter was most suitable for sulfide oxidation and growth of the cells.

• 社団法人日本生物工学会の論文
• 2001-05-25

著者

• Takahashl Mika

  Laboratory Of Animal Environment Meiji University

• Takahashi Masaya

  Industrial Chemistry The Kumamoto Institute Of Technology

• Kobayashi Shinya

  National Research Institute Of Brewing

• SHIBATA HIROOMI

  Laboratory of Animal Environment, Meiji University

• YAMAGUCHI IKUKO

  Laboratory of Animal Environment, Meiji University

• KOBAYASHI SHIGEKI

  Laboratory of Animal Environment, Meiji University

• Shibata Haruka

  Laboratory Of Animal Environment Meiji University

• Yamaguchi Ikuko

  Laboratory Of Animal Environment Meiji University

• Takahashi Mika

  Laboratory of Animal Environment, Meiji University

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