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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2017, Vol. 37 Issue (7): 64-71    DOI: 10.13523/j.cb.20170712
研究报告     
同源过表达BglR对嗜热毁丝霉β-葡萄糖苷酶活性的影响
来亚鹏1,2, 邓婷婷1,2, 刘刚1, 王娟1
1. 深圳大学生命与海洋科学学院 深圳市微生物基因工程重点实验室 深圳 518060;
2. 深圳市海洋生物资源与生态环境重点实验室 深圳 518060
The Influence of Homologous Overexpression of BglR on β-glucosidase Activities in Myceliophthora thermophila
LAI Ya-peng1,2, DENG Ting-ting1,2, LIU Gang1, WANG Juan1
1. College of Life Sciences and Oceanography, Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen University, Shenzhen 518060, China;
2. Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen 518060, China
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摘要: 目的:克隆嗜热毁丝霉(Myceliophthora thermophila ATCC42464)bglr基因,构建Mtbglr过表达载体,研究同源过表达bglr对嗜热毁丝霉β-葡萄糖苷酶活性及总纤维素酶活性的影响。方法:利用SLIC技术构建Mtbglr过表达载体;使用MtPpdc启动子及MtTpdc终止子将该基因进行同源过表达;利用原生质体转化、实时荧光定量PCR和酶活测定等技术实现bglr基因在嗜热毁丝霉中表达及酶活水平的鉴定。结果:成功构建Mtbglr过表达载体,并转化嗜热毁丝霉,结果表明,诱导培养条件下,转化子Mt8菌株的β-葡萄糖苷酶活性和胞外蛋白质浓度分别是WT菌株的1.7倍和1.9倍。结论: bglr基因对嗜热毁丝霉β-葡萄糖苷酶活性具有增强作用,为嗜热真菌β-葡萄糖苷酶基因调控奠定了理论基础。
关键词: 嗜热毁丝霉BglR嗜热真菌β-葡萄糖苷酶同源过表达    
Abstract: Objective: The influence of overexpression of β-glucosidase regulator BglR on beta-glucosidase activities in Myceliophthora thermophila ATCC42464 by cloning bglr gene and constructing Mtbglr overexpression vector were focused on. Methods: The SLIC was adopted to construct Mtbglr overexpression vector and the promoter of MtPpdc and terminator of MtTpdc was used for overexpress bglr gene. The gene expression and β-glucosidase activities were observed by protoplast transformation, real-time quantitative PCR and enzymatic determination. Results: The bglr gene was overexpressed in M. thermophila successfully. The result showed that the β-glucosidase activity and secreted protein concentration of transformant strain Mt8 were 1.7 and 1.9 fold higher, respectively, than that of wild type WT. Conclusion: The expression of bglr increased the β-glucosidase activity in M. thermophila under the inducing condition, which laid the theoretical foundation for the regulation of β-glucosidase gene of thermophilic fungi.
Key words: Homologous overexpression    β-glucosidase    Thermophilic fungi    BglR    M.thermophila
收稿日期: 2017-01-17 出版日期: 2017-07-25
ZTFLH:  Q815  
基金资助: 深圳市科技计划基础研究项目资助项目(JYJC20140418091413587)
通讯作者: 王娟     E-mail: wangjuan@szu.edu.cn
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引用本文:

来亚鹏, 邓婷婷, 刘刚, 王娟. 同源过表达BglR对嗜热毁丝霉β-葡萄糖苷酶活性的影响[J]. 中国生物工程杂志, 2017, 37(7): 64-71.

LAI Ya-peng, DENG Ting-ting, LIU Gang, WANG Juan. The Influence of Homologous Overexpression of BglR on β-glucosidase Activities in Myceliophthora thermophila. China Biotechnology, 2017, 37(7): 64-71.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170712        https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I7/64

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