Wine lees is a low value biomass resource rich in cellulose, with great potential for producing organic fertilizers and chemicals. However, the high acidity of wine lees limits the catalytic efficiency of the conversion tool endoglucanase. Here, we expressed endoglucanase tCel5A from Trichoderma reesei in Pichia pastoris, and the combination of promoter AOX1 and signal peptide SUC2 resulted in a highly active expression of 4632.81 U/mg. Subsequently, the catalytic center design and surface charge modification strategy resulted in mutants T88H/W255H and S45D/T55D/T59D exhibiting catalytic activity twice and three times higher than WT at pH 3.0, respectively. Finally, when the solid–liquid ratio was 1:15 (w/v), the degradation rate of wine lees was nearly double that of WT. The degradation products contained a variety of industrial and pharmaceutical raw components, including the antioxidant and anticonvulsant isopiperolein B. This study accelerates the green and sustainable management of wine lees.

中文翻译：

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内切葡聚糖酶降解酒糟的抗酸工程


酒糟是一种富含纤维素的低价值生物质资源，在生产有机肥料和化学品方面具有巨大潜力。然而，酒糟的高酸度限制了转化工具内切葡聚糖酶的催化效率。在这里，我们在毕赤酵母中表达来自里氏木霉的内切葡聚糖酶 tCel5A，启动子 AOX1 和信号肽 SUC2 的组合导致 4632.81 U/mg 的高活性表达。随后，催化中心设计和表面电荷修饰策略导致突变体 T88H/W255H 和 S45D/T55D/T59D 在 pH 3.0 时分别表现出比 WT 高 2 倍和 3 倍的催化活性。最后，当固液比为 1：15 （w/v） 时，酒糟的降解速率几乎是 WT 的两倍。降解产物含有多种工业和医药原料成分，包括抗氧化剂和抗惊厥药异哌霉菌素 B。本研究加速了酒糟的绿色和可持续管理。