• Title/Summary/Keyword: respiratory deficient transformant

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Improvement of Production and Secretion of Heterologous \alpha-Amylase from Saccharomyces cerevisiae. (외래 알파아밀라제의 Saccharomyces cerevisiae에서의 생산과 분비효율의 증진)

  • Choi, Sung-Ho;Kim, Geun
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.36-41
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    • 2003
  • In order to increase the production and secretion rate of mouse salivary $\alpha$-amylase from Saccharomyces cerevisiae, various experiments were attempted. A plasmid pCNNinv (AMY) was constructed by the substitution of ADCl promoter and native signal sequence of mouse salivary $\alpha$-amylase cDNA gene with PRBI promoter and yeast invertase leader sequence, which resulted in 25% increase in the production of $\alpha$-amylase in the culture medium. The respiratory deficient transformant carrying pCNNinv (AMY) were obtained by treating yeast cells with ethidium bromide, and the $\alpha$-amylase activities in the culture brothes of the respiratory-deficient transformants were 5-8 times higher than that of parental wild type strain. $\alpha$-Amylase activity was also increased 3 times when the 0.015% (w/v) of 2-mercaptoethanol was added to the culture medium.