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黑曲霉葡萄糖氧化酶在解脂耶氏酵母中的表达与特性分析

Expression and Characterization of Glucose Oxidase from Aspergillus niger in Yarrowia lipolytica.

作者信息

Khadivi Derakshan Fatemeh, Darvishi Farshad, Dezfulian Mehrouz, Madzak Catherine

机构信息

Department of Microbiology, Karaj Branch, Azad University, Karaj, Iran.

Microbial Biotechnology and Bioprocess Engineering (MBBE) Group, Department of Microbiology, Faculty of Science, University of Maragheh, Maragheh, Iran.

出版信息

Mol Biotechnol. 2017 Aug;59(8):307-314. doi: 10.1007/s12033-017-0017-8.

Abstract

Glucose oxidase (GOX) is currently used in clinical, pharmaceutical, food and chemical industries. The aim of this study was expression and characterization of Aspergillus niger glucose oxidase gene in the yeast Yarrowia lipolytica. For the first time, the GOX gene of A. niger was successfully expressed in Y. lipolytica using a mono-integrative vector containing strong hybrid promoter and secretion signal. The highest total glucose oxidase activity was 370 U/L after 7 days of cultivation. An innovative method was used to cell wall disruption in current study, and it could be recommended to use for efficiently cell wall disruption of Y. lipolytica. Optimum pH and temperature for recombinant GOX activity were 5.5 and 37 °C, respectively. A single band with a molecular weight of 80 kDa similar to the native and pure form of A. niger GOX was observed for the recombinant GOX in SDS-PAGE analysis. Y. lipolytica is a suitable and efficient eukaryotic expression system to production of recombinant GOX in compered with other yeast expression systems and could be used to production of pure form of GOX for industrial applications.

摘要

葡萄糖氧化酶(GOX)目前应用于临床、制药、食品和化学工业。本研究的目的是在解脂耶氏酵母中表达和鉴定黑曲霉葡萄糖氧化酶基因。首次使用含有强杂交启动子和分泌信号的单整合载体,在解脂耶氏酵母中成功表达了黑曲霉的GOX基因。培养7天后,总葡萄糖氧化酶活性最高为370 U/L。本研究采用了一种创新方法进行细胞壁破坏,该方法可推荐用于高效破坏解脂耶氏酵母的细胞壁。重组GOX活性的最适pH和温度分别为5.5和37℃。在SDS-PAGE分析中,重组GOX观察到一条分子量为80 kDa的单带,与黑曲霉GOX的天然纯形式相似。与其他酵母表达系统相比,解脂耶氏酵母是生产重组GOX的合适且高效的真核表达系统,可用于工业应用中生产纯形式的GOX。

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