CotA漆酶定向进化、降解黄曲霉毒素B1作用机理及动物解毒效果研究

国基评审专家1V1指导 中标率高出同行96.8%
结合最新热点,提供专业选题建议
深度指导申报书撰写,确保创新可行
指导项目中标800+,快速提高中标率
微信扫码咨询
中文摘要
黄曲霉毒素B1(AFB1)对动物和人具有极强的致畸、致癌、致突变性,如何通过生物酶催化生成低毒或无毒代谢产物,对提高饲料/食品安全和人类健康意义重大。本课题组从一株高效降解AFB1的地衣芽孢杆菌ANSB821基因组中克隆到编码CotA漆酶的基因,并将其转入到大肠杆菌中外源表达,重组表达的CotA漆酶能够直接氧化降解AFB1,分离纯化降解产物后,经质谱(UPLC-TOF/MS)和核磁(NMR)鉴定,CotA催化AFB1生成一对差向异构体AFQ1和epi-AFQ1。本项目拟利用定点突变技术和随机突变技术,对CotA漆酶进行定向进化,获得表达量和AFB1降解酶活均显著提高的突变体;系统评价两种产物的细胞毒性和致突变性,对CotA降解AFB1的作用机理进行深入研究;再通过体外模拟消化液和肉鸡黄曲霉毒素攻毒试验,验证CotA漆酶对动物的解毒效果;旨在开发一种具有实际应用价值的新型黄曲霉毒素解毒酶。
英文摘要
Aflatoxin B1 (AFB1) is a substance with strong teratogenicity, carcinogenicity and mutagenicity in animal and human body. How to degrade AFB1 to low toxic or nontoxic metabolites is great important and significant for ensuring feed/food safety and human health. The gene encoding laccase CotA with high activity in degrading AFB1 has been cloned from the genome of Bacillus licheniformis ANSB821 and transformed into Escherichia coli successfully for an efficient experession in our laboratory. Experimental results showed that recombinant CotA protein could directly catalyze and degrade AFB1 under normal physiological conditions. After isolation and purification of metabolites, a couple of epimeride, AFQ1 and epi-AFQ1 were identified from metabolites using mass spectrometry (UPLC-TOF/MS) and nuclear magnetic resonance (NMR). In this project, we will increase the enzyme acitivity and expression quantity further using the most advanced site-directed mutagenesis and random mutagenesis techniques. The degradation mechanism of AFB1 by CotA will be further explored via evaluating the cytotoxicity and mutagenicity of two products using cell experiment and Ames assay. Then the effectiveness of CotA protein in practical application will be assessed via using an in vivo digestion model and an animal experiment with broilers challenged with AFB1. We are expecting to obtain a new aflatoxin detoxification enzyme with great value in feed/food application.
期刊论文列表
专著列表
科研奖励列表
会议论文列表
专利列表
Degradation of zearalenone and aflatoxin B1 by Lac2 from Pleurotus pulmonarius in the presence of mediators
在介体存在下,来自侧耳的 Lac2 对玉米赤霉烯酮和黄曲霉毒素 B1 的降解。
DOI:10.1016/j.toxicon.2021.08.003
发表时间:2021-08-17
期刊:TOXICON
影响因子:2.8
作者:Song, Yanyi;Wang, Yanan;Zhao, Lihong
通讯作者:Zhao, Lihong
DOI:10.1016/j.foodchem.2020.126877
发表时间:2020-09-30
期刊:FOOD CHEMISTRY
影响因子:8.8
作者:Guo, Yongpeng;Qin, Xiaojuan;Zhao, Lihong
通讯作者:Zhao, Lihong
DOI:10.1016/j.foodcont.2022.109549
发表时间:2022-12
期刊:Food Control
影响因子:6
作者:Yongpeng Guo;Yanan Wang;Yu Tang;Qiugang Ma;C. Ji;Lihong Zhao
通讯作者:Yongpeng Guo;Yanan Wang;Yu Tang;Qiugang Ma;C. Ji;Lihong Zhao
DOI:10.3390/toxins13060401
发表时间:2021-06-04
期刊:Toxins
影响因子:4.2
作者:Ma J;Liu Y;Guo Y;Ma Q;Ji C;Zhao L
通讯作者:Zhao L
Enzymatic characterization and application of soybean hull peroxidase as an efficient and renewable biocatalyst for degradation of zearalenone.
大豆皮过氧化物酶作为玉米赤霉烯酮降解的高效可再生生物催化剂的酶学表征和应用。
DOI:10.1016/j.ijbiomac.2024.129664
发表时间:2024
期刊:International journal of biological macromolecules
影响因子:8.2
作者:Yongpeng Guo;Yu Tang;Liangyu Zhang;Yanrong Liu;Qiugang Ma;Lihong Zhao
通讯作者:Lihong Zhao
玉米赤霉烯酮降解酶的基因克隆、外源表达及解毒特性研究
- 批准号:31772637
- 项目类别:面上项目
- 资助金额:60.0万元
- 批准年份:2017
- 负责人:赵丽红
- 依托单位:
枯草芽孢杆菌ANSB01G对饲料中玉米赤霉烯酮生物降解机制的研究
- 批准号:31301981
- 项目类别:青年科学基金项目
- 资助金额:24.0万元
- 批准年份:2013
- 负责人:赵丽红
- 依托单位:
国内基金
海外基金
