Cloning and expression of Penicillium?minioluteum dextranase in Saccharomyces?cerevisiae and its exploitation as a reporter in the detection of mycotoxins

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• 作者：Xingmin Li ; Stefan H. Millson ; Raymond D. Coker and Ivor H. Evans
• 关键词：Dextranase expression ; Mycotoxin bioassay ; Penicillium minioluteum ; Saccharomyces cerevisiae
• 刊名：Biotechnology Letters
• 出版年：2006
• 出版时间：December, 2006
• 年：2006
• 卷：28
• 期：23
• 页码：1955-1964
• 全文大小：223 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

A dextranase gene from Penicillium minioluteum (strain IMI068219) has been cloned, sequenced and expressed in Saccharomyces cerevisiae via fusion of the DNA segment encoding the mature dextranase protein with α-factor signal sequence, and insertion into the GAL1–controlled expression vector pYES2/CT. Galactose-induced expression yielded extracellular dextranase activity of 0.63 units/ml and cell-associated dextranase activity of 0.48 units/ml, after 24 h incubation. The dextranase construct was introduced into a strain of S. cerevisiae expressing the human cytochrome P450 3A4 (CYP3A4) and the cognate reductase, which was then used to develop a microplate toxicity bioassay. Toxicity was signalled as inhibition of dextranase activity, assayed fluorimetrically. This novel bioassay was assessed using six economically significant mycotoxins.