Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Simultaneous Expression of Pseudomonas sp. Endo-1,4$\beta$-Glucanase and $\beta$-1,4=Glucisidase Gene in Escherichia coli and Saccharomyces cerevisiae

Pseudomonas sp. Endo-1,4-$\beta$-Glucanase와 $\beta$-1,4-Glucosidase 유전자의 대장균 및 효모에서의 동시 발현

  • 김양우 (경상대학교 식품공학과) ;
  • 전성식 (경상대학교 식품공학과) ;
  • 정영철 (진주전문대학 식품영양과) ;
  • 성낙계 (경상대학교 식품공학과)
  • Published : 1995.12.01
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Abstract

We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

Keywords

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