Development of an efficient protein expression system in the thermophilic fungus Myceliophthora thermophila

文献类型： 外文期刊

第一作者： Li, Jinyang

作者： Li, Jinyang;Wang, Yidi;Yang, Kun;Wang, Xiaolu;Wang, Yuan;Zhang, Honglian;Huang, Huoqing;Su, Xiaoyun;Yao, Bin;Luo, Huiying;Qin, Xing

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关键词： Myceliophthora thermophila; Protein expression; Promoter; 5'UTR; Signal peptide

期刊名称：MICROBIAL CELL FACTORIES （ 影响因子：6.4； 五年影响因子：6.3 ）

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年卷期： 2023 年 22 卷 1 期

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收录情况： SCI

摘要： Background Thermophilic fungus Myceliophthora thermophila has been widely used in industrial applications due to its ability to produce various enzymes. However, the lack of an efficient protein expression system has limited its biotechnological applications.Results In this study, using a laccase gene reporting system, we developed an efficient protein expression system in M. thermophila through the selection of strong constitutive promoters, 5'UTRs and signal peptides. The expression of the laccase was confirmed by enzyme activity assays. The results showed that the Mtpdc promoter (Ppdc) was able to drive high-level expression of the target protein in M. thermophila. Manipulation of the 5'UTR also has significant effects on protein expression and secretion. The best 5'UTR (NCA-7d) was identified. The transformant containing the laccase gene under the Mtpdc promoter, NCA-7d 5'UTR and its own signal peptide with the highest laccase activity (1708 U/L) was obtained. In addition, the expression system was stable and could be used for the production of various proteins, including homologous proteins like MtCbh-1, MtGh5-1, MtLPMO9B, and MtEpl1, as well as a glucoamylase from Trichoderma reesei.Conclusions An efficient protein expression system was established in M. thermophila for the production of various proteins. This study provides a valuable tool for protein production in M. thermophila and expands its potential for biotechnological applications.

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