Chen Xian-Zhong, Shen Wei, Fan You, Wang Zheng-Xiang
Jiangnan University, Wuxi, China.
Yi Chuan. 2011 Oct;33(10):1067-78. doi: 10.3724/sp.j.1005.2011.01067.
Filamentous fungi are used in a variety of industrial processes including the production of primary metabolites (e.g., organic acid, vitamins, and extracellular enzymes) and secondary metabolites (e.g., antibiotics, alkaloids, and gibberellins). Moreover, filamentous fungi have become preferred cell factories for production of foreign (heterologous) proteins in biotechnology in recent years. Compared to bacterial and yeast hosts, filamentous fungi showed predominant features such as the ability of growing on rather simple and inexpensive substrates, producing and secreting exceptionally large amounts of proteins, post-translational modifications, and GRAS (generally regarded as safe) approval. Therefore, the exploration of filamentous fungi has been attractive recently. This review summarizes the recent development in genomics, comparative genomics, transcriptomics, proteomics and metabolomics of filamentous fungi, and describes their applications and functions in reconstruction of metabolic network, discovery of novel proteins and genes, investigation of cell physiological and biochemical reactions, and strain breeding. This review also analyzes the bottlenecks of heterologous protein expression in filamentous fungi. Furthermore, special emphasis is given on the strategies for improving the protein production, including fusion expression of heterologous proteins, RNAi technology, manipulations of secretion pathways, codon optimization of foreign genes, and screening of protease mutants. Lastly, this review proposes the future direction of metabolic engineering of filamentous fungi.
丝状真菌被用于多种工业过程，包括初级代谢产物（如有机酸、维生素和胞外酶）和次级代谢产物（如抗生素、生物碱和赤霉素）的生产。此外，近年来丝状真菌已成为生物技术中生产外源（异源）蛋白的首选细胞工厂。与细菌和酵母宿主相比，丝状真菌具有显著特征，如能够在相当简单且廉价的底物上生长、产生和分泌大量蛋白质、进行翻译后修饰以及获得“一般认为安全”（GRAS）认证。因此，近年来对丝状真菌的探索备受关注。本综述总结了丝状真菌在基因组学、比较基因组学、转录组学、蛋白质组学和代谢组学方面的最新进展，并描述了它们在代谢网络重建、新蛋白质和基因发现、细胞生理生化反应研究以及菌株育种中的应用和功能。本综述还分析了丝状真菌中外源蛋白表达的瓶颈。此外重点介绍了提高蛋白质产量的策略，包括异源蛋白的融合表达、RNA干扰技术、分泌途径的操控、外源基因的密码子优化以及蛋白酶突变体的筛选。最后，本综述提出了丝状真菌代谢工程的未来发展方向。
