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首页> 外文期刊>Applied Microbiology >Podospora anserina Hemicellulases Potentiate the Trichoderma reesei Secretome for Saccharification of Lignocellulosic Biomass
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Podospora anserina Hemicellulases Potentiate the Trichoderma reesei Secretome for Saccharification of Lignocellulosic Biomass

机译：Podospora anserina半纤维素酶可增强里氏木霉分泌蛋白用于木质纤维素生物质糖化的能力

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摘要

To improve the enzymatic hydrolysis (saccharification) of lignocellulosic biomass by Trichoderma reesei , a set of genes encoding putative polysaccharide-degrading enzymes were selected from the coprophilic fungus Podospora anserina using comparative genomics. Five hemicellulase-encoding genes were successfully cloned and expressed as secreted functional proteins in the yeast Pichia pastoris . These novel fungal CAZymes belonging to different glycoside hydrolase families ( Pa Man5A and Pa Man26A mannanases, Pa Xyn11A xylanase, and Pa Abf51A and Pa Abf62A arabinofuranosidases) were able to break down their predicted cognate substrates. Although Pa Man5A and Pa Man26A displayed similar specificities toward a range of mannan substrates, they differed in their end products, suggesting differences in substrate binding. The N-terminal CBM35 module of Pa Man26A displayed dual binding specificity toward xylan and mannan. Pa Xyn11A harboring a C-terminal CBM1 module efficiently degraded wheat arabinoxylan, releasing mainly xylobiose as end product. Pa Abf51A and Pa Abf62A arabinose-debranching enzymes exhibited differences in activity toward arabinose-containing substrates. Further investigation of the contribution made by each P. anserina auxiliary enzyme to the saccharification of wheat straw and spruce demonstrated that the endo-acting hemicellulases ( Pa Xyn11A, Pa Man5A, and Pa Man26A) individually supplemented the secretome of the industrial T. reesei CL847 strain. The most striking effect was obtained with Pa Man5A that improved the release of total sugars by 28% and of glucose by 18%, using spruce as lignocellulosic substrate.
机译：为了改善里氏木霉对木质纤维素生物质的酶促水解（糖化作用），使用比较基因组学，从共亲真菌波氏假单胞菌中选择了一组编码假定的多糖降解酶的编码基因。成功地克隆了五个半纤维素酶编码基因，并在酵母毕赤酵母中表达为分泌的功能蛋白。这些新颖的真菌CAZymes属于不同的糖苷水解酶家族（Pa Man5A和Pa Man26A甘露聚糖酶，Pa Xyn11A木聚糖酶，Pa Abf51A和Pa Abf62A阿拉伯呋喃糖苷酶）能够分解其预测的同源底物。尽管Pa Man5A和Pa Man26A对多种甘露聚糖底物显示出相似的特异性，但它们的终产物不同，表明底物结合力不同。 Pa Man26A的N端CBM35模块显示了对木聚糖和甘露聚糖的双重结合特异性。带有C端CBM1模块的Pa Xyn11A有效降解了小麦阿拉伯木聚糖，主要释放木糖为最终产物。 Pa Abf51A和Pa Abf62A阿拉伯糖脱支酶对含阿拉伯糖的底物表现出不同的活性。进一步研究每种鹅膏假单胞菌辅助酶对小麦秸秆和云杉糖化的贡献，表明内用半纤维素酶（Pa Xyn11A，Pa Man5A和Pa Man26A）分别补充了里氏木霉CL847的分泌组。应变。使用云杉作为木质纤维素底物，使用Pa Man5A可获得最显着的效果，使总糖的释放提高了28％，葡萄糖的释放提高了18％。

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