过氧化物酶A4-Prx在毕赤酵母中的优化表达  

Fermentation Optimization of a Novel Recombinant Peroxiredoxin in Pichia pastoris for Zearalenone Detoxification

作　　者：陈艺[1] 朱珍[2] 吴晖[1] 钟凤[1] 唐语谦[1] 赖富饶[1] 

机构地区：[1]华南理工大学轻工与食品学院,广东广州510640 [2]广州市轻工职业学校,广东广州510650

出　　处：《现代食品科技》2014年第5期209-217,155,共10页Modern Food Science and Technology

基　　金：国家自然科学基金(31201330);广州市科技攻关项目(201300000202);中央高校基本科研业务费项目(2013ZZ0077;2013ZM0065);粮食公益性行业科研专项(201313005)

摘　　要：以来源于不动杆菌Acinetobacter sp.SM04的过氧化物酶A4-Prx的毕赤酵母工程表达菌株GS115/pPIC9K-A4-Prx为研究对象,优化其表达培养条件以提高该菌株对于目的蛋白A4-Prx的表达量。本论文首先研究了培养基成分与诱导条件对表达量的影响,结果表明培养基的pH、甘氨酸浓度和诱导温度对外源蛋白的产量均有显著影响。采用Box-Behnken设计,利用Design Expert软件进行二次回归分析得到了目的蛋白的最优表达条件为:诱导培养基pH 7.0、甘氨酸浓度为0.11%及诱导温度30℃。在此优化条件下重组蛋白的理论表达量达129.87 mg/L,约为未优化下的2倍,实验验证实际表达量达128.94 mg/L,且重组表达的过氧化物酶A4-Prx对酒糟蛋白饲料(DDGS)和食品中的玉米赤霉烯酮毒素(Zearalenone,ZEA)具有高效降解能力。本研究为过氧化物酶的工业化高密度发酵奠定了基础,推动生物降解ZEA研究的进展。In order to maximize the expression of a novel peroxiredoxin purified from Acinetobacter sp.SM04 in recombinant Pichia pastoris GS115/pPIC9K-A4-Prx,a single-factor test coupled with response surface methodology（RSM） was used to adjust the induction condition.The results showed that pH value,temperature and glycine concentration were significant influencing factors for A4-Prx expression,and the optimized induction conditions were as follows：pH 7,0.11% glycine at 30 ℃,and the theoretic yield of A4-Prx could reach to 129.873 mg/L.Then further experiments under the optimized induction conditions illustrated that the concentration of the novel peroxiredoxin synthesized by Pichia pastoris GS115/pPIC9K-A4-Prx was 128.941 mg/L and had a significantly ZEA-degrading effect on DDGS and some food materials.This study laid the foundation for the high-density industrial fermentation of peroxiredoxin as well as the development of ZEA degradation.

关 键 词：过氧化物酶 毕赤酵母 玉米赤霉烯酮 响应面分析 生物降解 

分 类 号：TQ925[轻工技术与工程—发酵工程]