title: Enhanced cellulase production from Trichoderma reesei Rut-C30 by engineering with an artificial zinc finger protein library.
abstract: Trichoderma reesei Rut-C30 is a well-known cellulase producer, and improvement of its cellulase production is of great interest. An artificial zinc finger protein (AZFP) library is constructed for expression in T. reesei Rut-C30, and a mutant strain T. reesei U3 is selected based on its enhanced cellulase production. The U3 mutant shows a 55% rise in filter paper activity and 8.1-fold increased β-glucosidase activity, when compared to the native strain T. reesei Rut-C30. It is demonstrated that enhanced β-glucosidase activity was due to elevated transcription level of β-glucosidase gene in the U3 mutant. Moreover, significant elevation in transcription levels of several putative Azfp-U3 target genes is detected in the U3 mutant, including genes encoding hypothetical transcription factors and a putative glycoside hydrolase. Furthermore, U3 cellulase shows 115% higher glucose yield from pretreated corn stover, when compared to the cellulase of T. reesei Rut-C30. These results demonstrate that AZFP can be used to improve cellulase production in T. reesei Rut-C30. Our current work offers the establishment of an alternative strategy to develop fungal cell factories for improved production of high value industrial products.
authors: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, China.
authors: Fei
authors: F
authors: Zhang
authors: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, China.|State Key Laboratory of Microbial Metabolism and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
authors: Fengwu
authors: F
authors: Bai
authors: State Key Laboratory of Microbial Metabolism and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, China. xqzhao@sjtu.edu.cn.
authors: Xinqing
authors: X
authors: Zhao
mesh_terms: D002480:Cellulase / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D009154:Mutation; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: D011994:Recombinant Proteins; D014157:Transcription Factors; D002480:Cellulase
keywords: Artificial transcription factor (ATF); Artificial zinc finger protein (AZFP); Cellulase; Trichoderma reesei Rut-C30; β-glucosidase