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A sensitive bioassay for the mycotoxin aflatoxin B1, which also responds to the mycotoxins aflatoxin G1 and T-2 toxin, using engineered baker's yeast

A sensitive bioassay for the mycotoxin aflatoxin B1, which also responds to the mycotoxins aflatoxin G1 and T-2 toxin, using engineered baker's yeast

Li, Xingmin, Millson, Stefan, Coker, Raymond and Evans, Ivor (2009) A sensitive bioassay for the mycotoxin aflatoxin B1, which also responds to the mycotoxins aflatoxin G1 and T-2 toxin, using engineered baker's yeast. Journal of Microbiological Methods, 77 (3). pp. 285-291. ISSN 0167-7012 (doi:https://doi.org/10.1016/j.mimet.2009.03.003)

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Abstract

A novel aflatoxin B(1) bioassay was created by introducing a Lipomyces kononenkoae alpha-amylase gene into a strain of S. cerevisiae capable of expressing the human cytochrome P450 3A4 (CYP3A4), and the cognate human CYP450 reductase. This strain and a dextranase-expressing strain were used in the development of a microtitre plate mycotoxin bioassay, which employed methanol as the solvent and polymyxin B nonapeptide as a permeation enhancer. Stable co-expression of the CYP3A4 gene system and of the dextranase and amylase genes in the two bioassay strains was demonstrated. The bioassay signalled toxicity as inhibition of secreted carbohydrase activity, using sensitive fluorimetric assays. The amylase-expressing strain could detect aflatoxin B(1) at 2 ng/ml, and was more sensitive than the dextranase-expressing strain. Aflatoxin G(1) could be detected at 2 microg/ml, and the trichothecene mycotoxin T-2 toxin was detectable at 100 ng/ml.

Item Type: Article
Uncontrolled Keywords: mycotoxin, bioassay, yeast, aflatoxin B1, alpha-amylase
Subjects: R Medicine > RB Pathology
Q Science > QD Chemistry
Faculty / School / Research Centre / Research Group: Faculty of Education, Health & Human Sciences > School of Human Sciences (HUM)
Related URLs:
Last Modified: 09 Oct 2021 04:46
URI: http://gala.gre.ac.uk/id/eprint/1586

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