Synthetic multi-component enzyme mixtures for deconstruction of lignocellulosic biomass
Synthetic multi-component enzyme mixtures for deconstruction of lignocellulosic biomass
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DOI：
10.1016/j.biortech.2010.07.028
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发表时间：
2010-12-01
影响因子：
11.4
通讯作者：
Walton, Jonathan D.
中科院分区：
文献类型：
--
作者：
Banerjee, Goutami;Car, Suzana;Walton, Jonathan D.
使用称为GENPLAT的高通量酶测定平台来指导来自10种“辅助”酶和6种“核心”酶的单个纯化酶的优化混合物的开发。优化酶混合物用于从氨纤维膨胀（AFEX）预处理的玉米秸秆中释放Glu、Xyl或两者的组合。测定条件为15 mg/g葡聚糖的固定酶载量、48 h消化和50 ℃。与单独的核心组相比，十种测试的辅助蛋白中的五种增强了Glu或Xyl产率，而五种没有。含有核心组和针对谷氨酸优化的五种辅助酶的11种组分混合物释放了52.1%的可用谷氨酸，而单独使用核心组的释放率为38.5%。针对Xyl优化的混合物释放了39.9%的Xyl，相比之下，单独的核心组释放了26.4%。我们预测，仍有相当大的机会，进一步改善合成混合物。此外，本文所述的策略适用于开发用于任何预处理/生物质组合的更有效的酶混合物，以及用于检测对木质纤维素解构做出迄今为止未被认识到的贡献的酶。(C)2010爱思唯尔有限公司版权所有。
A high throughput enzyme assay platform, called GENPLAT, was used to guide the development of an optimized mixture of individual purified enzymes from ten "accessory" and six "core" enzymes. Enzyme mixtures were optimized for release of Glu, Xyl, or a combination of the two from corn stover pretreated by ammonia-fiber expansion (AFEX). Assay conditions were a fixed enzyme loading of 15 mg/g glucan, 48 h digestion, and 50 degrees C. Five of the ten tested accessory proteins enhanced Glu or Xyl yield compared to the core set alone, and five did not. An 11-component mixture containing the core set and five accessory enzymes optimized for Glu released 52.1% of the available Glu, compared to 38.5% with the core set alone. A mixture optimized for Xyl released 39.9% of the Xyl, compared to 26.4% with the core set alone. We predict that there is still considerable opportunity for further improvement of synthetic mixtures. Furthermore, the strategy described here is applicable to the development of more efficient enzyme cocktails for any pretreatment/biomass combination and for detecting enzymes that make a heretofore unrecognized contribution to lignocellulose deconstruction. (C) 2010 Elsevier Ltd. All rights reserved.