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[Artificial zinc finger protein mediated cellulase production in Trichoderma reesei Rut-C30].
Sheng Wu Gong Cheng Xue Bao. 2019 Jan 25; 35(1):81-90.SW

Abstract

Trichoderma reesei Rut-C30 is widely used in industrial cellulase production, and development of cellulase hyper-producer is of great importance for economic lignocellulosic biorefinery. In this study, T. reesei Rut-C30 was engineered with an artificial zinc finger proteins (AZFPs) library. Two mutants T. reesei M1 and M2 with improved cellulase production were obtained. Compared to the parent strain, the filter paper activity (FPase) of T. reesei M1 and M2 increased 100% and 53%, respectively. In addition, the total amount of extracellular protein from the M1 mutant increased 69%, whereas the endo-β-glucanase (CMCase) activity of the M2 mutant is 64% higher compared to the parental strain. Furthermore, RT-qPCR analysis showed that the major cellulase genes exhibited significantly increased expression in both mutants, but different patterns were observed in the two mutants. On the other hand, the cellulase transcriptional repressor ace1 was down-regulated in both mutants, but the transcription level of the activator xyr1 was only up-regulated in the strain M1. These results demonstrated that different AZFPs exert diverse regulatory mechanisms on cellulase production in T. reesei. Analysis of the target genes of AZFPs from T. reesei M1 and M2 will not only benefit further exploration of the regulatory mechanisms of cellulase biosynthesis in T. reesei, but also enable development of cellulase hyper-producing strains by metabolic engineering.

Authors+Show Affiliations

School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, Liaoning, China.State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China. Joint International Research Laboratory of Metabolic & Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, China.State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China. Joint International Research Laboratory of Metabolic & Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, China.

Pub Type(s)

Journal Article

Language

chi

PubMed ID

30756537

Citation

Meng, Qingshan, et al. "[Artificial Zinc Finger Protein Mediated Cellulase Production in Trichoderma Reesei Rut-C30]." Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, vol. 35, no. 1, 2019, pp. 81-90.
Meng Q, Li J, Zhang F, et al. [Artificial zinc finger protein mediated cellulase production in Trichoderma reesei Rut-C30]. Sheng Wu Gong Cheng Xue Bao. 2019;35(1):81-90.
Meng, Q., Li, J., Zhang, F., Zhao, X., & Bai, F. (2019). [Artificial zinc finger protein mediated cellulase production in Trichoderma reesei Rut-C30]. Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, 35(1), 81-90. https://doi.org/10.13345/j.cjb.180132
Meng Q, et al. [Artificial Zinc Finger Protein Mediated Cellulase Production in Trichoderma Reesei Rut-C30]. Sheng Wu Gong Cheng Xue Bao. 2019 Jan 25;35(1):81-90. PubMed PMID: 30756537.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Artificial zinc finger protein mediated cellulase production in Trichoderma reesei Rut-C30]. AU - Meng,Qingshan, AU - Li,Jiaxiang, AU - Zhang,Fei, AU - Zhao,Xinqing, AU - Bai,Fengwu, PY - 2019/2/14/entrez PY - 2019/2/14/pubmed PY - 2019/6/14/medline KW - Trichoderma reesei KW - artificial zinc finger protein KW - cellulase KW - metabolic engineering KW - transcription factor SP - 81 EP - 90 JF - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JO - Sheng Wu Gong Cheng Xue Bao VL - 35 IS - 1 N2 - Trichoderma reesei Rut-C30 is widely used in industrial cellulase production, and development of cellulase hyper-producer is of great importance for economic lignocellulosic biorefinery. In this study, T. reesei Rut-C30 was engineered with an artificial zinc finger proteins (AZFPs) library. Two mutants T. reesei M1 and M2 with improved cellulase production were obtained. Compared to the parent strain, the filter paper activity (FPase) of T. reesei M1 and M2 increased 100% and 53%, respectively. In addition, the total amount of extracellular protein from the M1 mutant increased 69%, whereas the endo-β-glucanase (CMCase) activity of the M2 mutant is 64% higher compared to the parental strain. Furthermore, RT-qPCR analysis showed that the major cellulase genes exhibited significantly increased expression in both mutants, but different patterns were observed in the two mutants. On the other hand, the cellulase transcriptional repressor ace1 was down-regulated in both mutants, but the transcription level of the activator xyr1 was only up-regulated in the strain M1. These results demonstrated that different AZFPs exert diverse regulatory mechanisms on cellulase production in T. reesei. Analysis of the target genes of AZFPs from T. reesei M1 and M2 will not only benefit further exploration of the regulatory mechanisms of cellulase biosynthesis in T. reesei, but also enable development of cellulase hyper-producing strains by metabolic engineering. SN - 1000-3061 UR - https://www.unboundmedicine.com/medline/citation/30756537/[Artificial_zinc_finger_protein_mediated_cellulase_production_in_Trichoderma_reesei_Rut_C30]_ L2 - https://doi.org/10.13345/j.cjb.180132 DB - PRIME DP - Unbound Medicine ER -
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