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Microbiology and Biotechnology Letters

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Microbiol. Biotechnol. Lett. 1998; 26(5): 406-412

Saccharomyces cerevisiae에서 Trichoderma Endoglucanase의 발현과 분비

Expression and Secretion of Trichodema Endoglucanase in Saccharomyces cerevisiae.

Abstract

The endoglucanase gene, egl6, of Trichoderma sp. was connected with the yeast ADH1 promoter, and the resultant plasmid, pVT-C4, was introduced into three S. cerevisiae host strains (YNN27, 2805, and SEY2102). Among each 80 transformants, the cell growth and expression level of endoglucanase were compared in test-tube cultivation, and three respective transformants for each host cells showing the highest expression level and cell growth were selected. When three recombinant yeast cells were batchwise cultivated for 48 hr in flask, the total activities of endoglucanase expressed were about 1140 unit/l with 2805/pVT-C4, 1020 unit/l with SEY2102/pVT-C4, and 590 unit/l with YNN27/pVT-C4. Irrespective of host strain, about 80% of the expressed endoglucanase was detected in the extracellular medium. In addition, it was also found that the recombinant enzyme was secreted into the culture medium as two major forms of lightly and heavily glycosylated proteins.

Keywords: Saccharomyces cerevisiae, expression, Trichoderma endoglucanase, ADH1 promoter, glycosylation

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