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

Li, Xingmin and Millson, Stefan H. and Coker, Raymond D. and Evans, Ivor H. (2006) Cloning and expression of Penicillium minioluteum dextranase in Saccharomyces cerevisiae and its exploitation as a reporter in the detection of mycotoxins. Biotechnology Letters, 28 . pp. 1955-1964. ISSN 0141-5492

Full content URL: http://dx.doi.org/10.1007/s10529-006-9183-7

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Item Type:Article
Item Status:Live Archive

Abstract

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.

Keywords:Dextranase expression, Mycotoxin bioassay, Penicillium minioluteum, Saccharomyces cerevisiae
Subjects:C Biological Sciences > C560 Biotechnology
C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
C Biological Sciences > C500 Microbiology
Divisions:College of Science > School of Life Sciences
ID Code:15327
Deposited On:07 Oct 2014 17:31

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