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 (Print), 1872-8359 (Online) (doi:10.1016/j.mimet.2009.03.003)

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Abstract

A novel aflatoxin B1 bioassay was created by introducing a Lipomyces kononenkoae α-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 B1 at 2 ng/ml, and was more sensitive than the dextranase-expressing strain. Aflatoxin G1 could be detected at 2 µg/ml, and the trichothecene mycotoxin T-2 toxin was detectable at 100 ng/ml.

Item Type:	Article
Additional Information:	** Article version: AM ** Embargo end date: 31-12-9999 ** From Elsevier via Jisc Publications Router ** History: accepted 08-03-2009; issue date 30-06-2009. ** Licence for AM version of this article: This article is under embargo with an end date yet to be finalised.
Uncontrolled Keywords:	mycotoxin; bioassay; yeast; aflatoxin B1; alpha-amylase
Subjects:	Q Science > Q Science (General) S Agriculture > S Agriculture (General) T Technology > T Technology (General)
Pre-2014 Departments:	School of Science
SWORD Depositor:	Users 6393 not found.
Last Modified:	27 May 2024 14:54
URI:	http://gala.gre.ac.uk/id/eprint/32278

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