摘要
【背景】玉米赤霉烯酮(zearalenone,ZEN)及其衍生物是一群具有雌激素活性的霉菌毒素,广泛存在于被霉菌污染的谷物中,造成食品业和畜牧业的巨大损失。利用专一性高的水解酶进行生物转化可有效去除玉米赤霉烯酮。【目的】构建高效表达玉米赤霉烯酮水解酶的酵母系统,以促进玉米赤霉烯酮水解酶的研究和工业应用。【方法】将来源于麦氏喙枝孢霉(RhinocladiellamackenzieiCBS650.93)的Rmzhd基因转入毕赤酵母中,筛选获得高效表达菌株,通过高效液相色谱分析发酵液中重组酶的性质。【结果】发酵液中Rm ZHD对ZEN的酶活力为16.67 U/m L,对α-ZOL的酶活力为9.85 U/m L。SDS-PAGE检测表达产物的分子量,与理论值30.7k D符合,且发酵上清液蛋白纯度高。Rm ZHD的最适p H值为9.6,最适温度为45°C,并具有较好的耐热性。【结论】研究结果为玉米赤霉烯酮水解酶的异源表达及其潜在的工业应用提供了一定的指导。
[Background]Zearalenone(ZEN)and its derivatives are a series of mycotoxins with estrogenic activity which are widespread in mold contaminated cereals.Mycotoxin contamination has caused huge loss to food and feed industries.To develop an efficient solution is an urgent task.[Objective]In order to promote the research and application of zearalenone hydrolase,we constructed the recombinant plasmid that highly expressed zearalenone hydrolase in Pichia pastoris.[Methods]The Rmzhd gene from Rhinocladiella mackenziei CBS 650.93 was transformed into Pichia pastoris GS115 for heterologous expression.The high expression strain was screened and the characterization of recombinant Rm ZHD was analyzed by high performance liquid chromatography(HPLC).[Results]The ZEN andα-ZOL degradation activities of the recombinant Rm ZHD from the supernatant were 16.67 U/m L and 9.85 U/m L,respectively.The molecular mass of Rm ZHD was about 30.7 k D,and the purity of protein was very high,as shown by SDS-PAGE.The optimal p H and temperature of recombinant Rm ZHD were determined as p H 9.6 and 45°C,and Rm ZHD has a good thermostability.[Conclusion]The results can provide guidance for the heterologous expression and potential industrial application of zearalenone hydrolase.
作者
刘文婷
梁爱玲
刘卫东
商娜
郭瑞庭
张同存
郑迎迎
LIU Wen-Ting;LIANG Ai-Ling;LIU Wei-Dong;SHANG-Na;GUO Rey-Ting;ZHANG Tong-Cun;ZHENG Ying-Ying(College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China;Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China)
出处
《微生物学通报》
CAS
CSCD
北大核心
2020年第7期2012-2020,共9页
Microbiology China
基金
国家重点研发计划(S2016G4513)。
