A kind of zearalenone detoxification method
A technology of zearalenone and lt-omt, applied in the field of zearalenone detoxification, can solve the problems of unstable enzyme activity, restrict the development of ZEN biological detoxification, toxic effects, etc., and achieve high detoxification efficiency , good detoxification effect
Active Publication Date: 2022-01-25
YANGZHOU UNIV
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Problems solved by technology
However, this technology also has shortcomings, such as the screened microorganisms are also animal pathogens; the secondary metabolites produced by microorganisms have toxic effects on animals; the intermediate products of ZEN degraded by biological enzymes still have toxic effects on animals; the yield of biological enzymes is low , complex separation and purification process, unstable enzyme activity, harsh enzyme action conditions, etc. These shortcomings also limit the development of ZEN biological detoxification field
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[0049] The present invention will be further described below in combination with specific embodiments and accompanying drawings.
[0050] (1). Experimental materials:
[0051] ① Strain and carrier:
[0052] E.coli DH5α competent cells were purchased from Kangwei Century Company;
[0053] BJ5464-NpgA: from the Institute of Biotechnology, Chinese Academy of Agricultural Sciences; PRS42 carrier: constructed by the Biosynthesis Laboratory of the Institute of Biotechnology, Chinese Academy of Agricultural Sciences.
[0054] ② Enzymes and kits:
[0055] Nde I, Pme I, T4 ligase, purchased from NEB company products;
[0056] Taq enzyme was purchased from Quanshijin Company;
[0057] Plasmid extraction and gel recovery kits were purchased from TIGEN Company and AXYGEN Company respectively.
[0058] Other experimental methods not specified in the examples were carried out according to conventional methods, and molecular cloning was carried out according to the conditions described ...
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Abstract
The invention belongs to the field of biotechnology, and relates to a detoxification method for zearalenone. The gene of Lt-OMT is connected with the pRS425 carrier to construct a recombinant artificial plasmid 92-Lt-OMT-YEp-Leu, and the artificial plasmid 92-Lt-YEp-Leu is constructed. OMT‑YEp‑Leu is transferred into competent yeast cells, the yeast cells are placed in Leu auxotrophic medium for culture, zearalenone ZEN is added to the Leu auxotrophic medium, and Lt‑OMT synthesized by yeast cells Conversion of Zearalenone ZEN to 3‑OCH 3 -ZEN. ZEN of the present invention is converted to 3-OCH by Lt-OMT 3 ‑ZEN, experimentally proven 3‑OCH 3 ‑ZEN has no obvious side effects on mice. In the yeast fermentation system, the conversion efficiency of Lt‑OMT to ZEN was over 98%, and there was almost no substrate residue in HPLC analysis.
Description
technical field [0001] The invention belongs to the field of biotechnology, and relates to a detoxification method of Zearalenone (Zearalenone, ZEN), which uses a Lasiodiplodia theobromae Oxymethyltransferase (Lasiodiplodia theobromae Oxymethyltransferase, Lt -OMT) converts the 3-hydroxyl (-OH) of zearalenone ZEN into an oxymethyl group (-OCH 3 ). Background technique [0002] Zearalenone (ZEN) is mainly Fusarium graminearum, F. tricinctum, F. culmorum, F. equiseti, Fusarium half-naked (F.sernitectum), Fusarium solanisis (F.solani) and other fungi produce secondary metabolites, widely present in grains and their products. Zearalenone ZEN has estrogen-like effects and can bind to estrogen receptors in animals to induce reproductive toxicity, teratogenicity, and carcinogenicity, and seriously affect animal reproductive performance. In addition, zearalenone ZEN can cause immune Inhibition, liver and kidney damage, digestion and absorption disorders and other hazards. [000...
Claims
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IPC IPC(8): C12P17/08C12N15/52C12N15/81C12N9/00C12N1/19
CPCC12N9/00C12P17/08
Inventor 李金贵刘明江时从来韩一晴徐玉泉
Owner YANGZHOU UNIV