期刊
BIOTECHNOLOGY JOURNAL
卷 11, 期 10, 页码 1282-1290出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201600227
关键词
Artificial zinc finger protein (AZFP); Cellulase; beta-glucosidase; Lignocellulosic biomass; Trichoderma reesei Rut-C30
资金
- National Research Foundation of China [31461143029, 5151101168]
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 beta-glucosidase activity, when compared to the native strain T. reesei Rut-C30. It is demonstrated that enhanced beta-glucosidase activity was due to elevated transcription level of beta-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.
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