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Exploring the Synergy between Cellobiose Dehydrogenase from Phanerochaete chrysosporium and Cellulase from Trichoderma reesei

机译:探索Phanerochaete chrysosporium的纤维二糖脱氢酶和里氏木霉的纤维素酶之间的协同作用

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摘要

Recent demands for the production of lignocellulose biofuels boosted research on cellulase. Hydrolysis efficiency and production cost of cellulase are two bottlenecks in “biomass to biofuels” process. The Trichoderma cellulase mixture is one of the most commonly used enzymes for cellulosic hydrolysis. During hydrolytic process cellobiose accumulation causes feedback inhibition against most cellobiohydrolases and endoglucanases. In this study, we demonstrated the synergism effects between cellobiose dehydrogenase (CDH) and cellulase both in vitro and in vivo. The CDH from Phanerochaete chrysosporium was heterologously expressed in Pichia pastoris. Supplementation of the purified CDH in Trichoderma cellulase increased the cellulase activities. Especially β-glucosidase activity was increased by 30–100% varying at different time points. On the other hand, the cdh gene was heterologously expressed in Trichoderma reesei to explore the synergism between CDH and cellulases in vivo. The analyses of gene expression and enzymatic profiles of filter paper activity, carboxymethylcellulase (CMCase) and β-glucosidase show the increased cellulase activity and the enhanced cellulase production in the cdh-expressing strains. The results elucidate a possible mechanism for diminishing the cellobiose inhibition of cellulase by CDH. These findings provide a novel perspective to make more economic enzyme cocktails for commercial application or explore alternative strategies for generating cellulase-producing strains with higher efficiency.
机译:对生产木质纤维素生物燃料的最新需求促进了对纤维素酶的研究。纤维素酶的水解效率和生产成本是“从生物质到生物燃料”过程的两个瓶颈。木霉纤维素酶混合物是纤维素水解最常用的酶之一。在水解过程中,纤维二糖的积累引起对大多数纤维二糖水解酶和内切葡聚糖酶的反馈抑制。在这项研究中,我们证明了纤维二糖脱氢酶(CDH)和纤维素酶在体外和体内的协同作用。 Phanerochaete chrysosporium的CDH在巴斯德毕赤酵母中异源表达。在木霉属纤维素酶中补充纯化的CDH可增加纤维素酶活性。特别是在不同时间点,β-葡萄糖苷酶的活性增加了30-100%。另一方面,cdh基因在里氏木霉中异源表达,以探索体内CDH和纤维素酶之间的协同作用。对滤纸活性,羧甲基纤维素酶(CMCase)和β-葡萄糖苷酶的基因表达和酶学特征的分析表明,在表达cdh的菌株中纤维素酶活性增加,纤维素酶产生增加。结果阐明了减轻CDH对纤维素酶的纤维二糖抑制作用的可能机制。这些发现提供了一种新颖的观点,可以使酶混合物更经济地用于商业应用,或者探索产生高产纤维素酶菌株的替代策略。

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