Engineering Trichoderma reesei for Hyperproduction of Cellulases on Glucose to Efficiently Saccharify Pretreated Corncobs.
Engineering Trichoderma reesei for Hyperproduction of Cellulases on Glucose to Efficiently Saccharify Pretreated Corncobs.
复制标题
工程化里氏木霉以葡萄糖为基础过量生产纤维素酶，以有效糖化预处理的玉米芯。
DOI：
10.1021/acs.jafc.0c04663
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发表时间：
2020
影响因子：
6.1
通讯作者：
Liu Weifeng
中科院分区：
文献类型：
--
作者：
Zheng Fanglin;Yang Renfei;Cao Yanli;Zhang Weixin;Lv Xinxing;Meng Xiangfeng;Zhong Yaohua;Chen Guanjun;Zhou Qingxin;Liu Weifeng
丝状真菌里氏木霉（Trichoderma reesei）是一种纤维素酶产生菌，在工业上有着广泛的应用.在这里，我们描述了合理的工程公开可用的T。reeseiQM 9414菌株在葡萄糖上实现了纤维素酶的显著高水平生产。通过铜阻遏型启动子Ptcu 1过表达关键的纤维素酶调节因子XYR 1，首先实现了在分解代谢产物阻遏（CCR）的背景下的完全纤维素酶生产，同时消除了诱导糖用于酶生产的要求。T。reeseibgl 1基因进一步过表达以补偿其对葡萄糖的低β-葡萄糖苷酶活性。与在Avicel上培养的CCR释放的RUT-C30菌株相比，这种过表达导致FPase活性增加102%。此外，工程菌粗酶液对预处理后的玉米芯渣的糖化效率比阿维菌素（Avicel）上培养的RUT-C30粗酶液提高了85%。工程师T。因此，在进一步优化农业废物的有效糖化之后，reeseisstrain显示出作为递送商业纤维素酶的可行替代物的巨大潜力。
The filamentous fungusTrichoderma reesei(teleomorphHypocrea jecorina) is widely used as a cellulase producer in the industry. Herein, we describe the rational engineering of the publicly availableT. reeseiQM9414 strain to achieve a remarkable high-level production of cellulase on glucose. Overexpression of the key cellulase regulator XYR1 by the copper-repressible promoter Ptcu1was first implemented to achieve a full cellulase production in the context of catabolite repression (CCR) while eliminating the requirement of inducing sugars for enzyme production. TheT. reeseibgl1gene was further overexpressed to compensate for its low β-glucosidase activity on glucose. This overexpression resulted in a 102% increase in FPase activity compared with the CCR-released RUT-C30 strain cultured on Avicel. Moreover, the saccharification efficiency toward pretreated corncob residues by crude enzymes from the engineered strain on glucose increased by 85% compared with that treated by enzymes from RUT-C30 cultivated on Avicel. The engineeredT. reeseistrain thus shows great potential as a viable alternative to deliver commercial cellulases after further optimization for efficient saccharification of agricultural waste.