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嗜热毁丝霉在可溶性淀粉上的转录分析及调节因子 AmyR 在多糖降解中的作用。

Transcriptional analysis of Myceliophthora thermophila on soluble starch and role of regulator AmyR on polysaccharide degradation.

机构信息

College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University. Nanjing 211800, China.

Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences. Tianjin 300308, China.

出版信息

Bioresour Technol. 2018 Oct;265:558-562. doi: 10.1016/j.biortech.2018.05.086. Epub 2018 May 24.

Abstract

Thermophilic fungus Myceliophthora thermophila has great capacity for biomass degradation and is an attractive option for use as cell factory to produce chemicals directly from renewable polysaccharides, such as starch, rather than monomer glucose. To date, there has been no transcriptomic analysis of this thermophilic fungus on starch. This study determined the transcriptomic profile of M. thermophila responding to soluble starch and a 342-gene set was identified as a "starch regulon", including the major amylolytic enzyme (Mycth_72393). Its overexpression led to increased amylase activities on starch by 35%. Furthermore, overexpressing the key amylolytic enzyme regulator AmyR in M. thermophila significantly increased amylase activity by 30%. Deletion of amyR by the CRISPR/Cas9 system led to the relief of carbon catabolite repression and 3-fold increased lignocellulase activities on cellulose. This study will accelerate rational fungal strain engineering for biochemical production from biomass substrates such as raw corn starch and even crop straw.

摘要

嗜热真菌嗜热毁丝霉具有很强的生物质降解能力,是一种很有吸引力的选择,可用作细胞工厂,直接从可再生多糖(如淀粉)而不是单体葡萄糖生产化学品。迄今为止,尚未对这种嗜热真菌在淀粉上的转录组进行分析。本研究确定了嗜热毁丝霉响应可溶性淀粉的转录组图谱,鉴定出一个包含主要淀粉分解酶(Mycth_72393)的 342 个基因集为“淀粉调控子”。其过表达使淀粉的淀粉酶活性提高了 35%。此外,在嗜热毁丝霉中过表达关键的淀粉分解酶调节因子 AmyR 可使淀粉酶活性显著提高 30%。通过 CRISPR/Cas9 系统敲除 amyR 可解除碳分解代谢物阻遏作用,使纤维素上的木质纤维素酶活性提高 3 倍。这项研究将加速从生物质底物(如原玉米淀粉甚至作物秸秆)进行生物化学生产的理性真菌菌株工程。

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