Engineering of a High-Throughput Screening System to Identify Cellulosic Biomass, Pretreatments, and Enzyme Formulations That Enhance Sugar Release
Engineering of a High-Throughput Screening System to Identify Cellulosic Biomass, Pretreatments, and Enzyme Formulations That Enhance Sugar Release
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DOI：
10.1002/bit.22527
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发表时间：
2010-02-01
影响因子：
3.8
通讯作者：
Wyman, Charles E.
中科院分区：
文献类型：
--
作者：
Studer, Michael H.;DeMartini, Jaclyn D.;Wyman, Charles E.
纤维素生物质是制造液体燃料的唯一丰富、可持续的原料，其顽固性是低成本生物加工的主要障碍，开发更容易转化的植物和更有效的酶将是非常有益的。因为没有单一的参数描述顽固性，所以只能通过测量经过预处理和酶解的植物的糖释放来识别更好的变种。然而，植物的基因改造加上解构蛋白的分子工程和预处理条件的定义创建了非常大的样本集，以前的生物质高温高压预处理方法阻止了完全集成的高通量(HTP)筛选管道的使用。在此，我们报告了一种新型HTP预处理系统的工程设计，该系统采用96孔板格式，可承受极端的预处理条件，用于快速筛选生物质-酶-预处理组合。这包括开发蒸汽加热和水淬火的新方法，使系统产生比以前可能的更快的热唇和冷小丑，并显示整个多孔板一致的温度历史。实验结果表明，该装置中多个孔板的预处理与酶解耦合性能与常规管式反应器的性能基本一致。生物技术。比昂斯。2010；105：231-238。(C)2009年威利期刊公司。
The recalcitrance Of Cellulosic biomass, the only abundant, sustainable feedstock for making liquid fuels, is a primary obstacle to low cost biological processing, and development of more easily converted plants and more effective enzymes would be of great benefit. Because no single parameter describes recalcitrance, Superior variants call only be identified by measuring sugar release from plants Subjected to pretreatment and enzymatic hydrolysis. However, genetic modifications of plants coupled with molecular engineering of deconstruction proteins and definition of pretreatment conditions create a very large sample set, and previous methods for biomass pretreatment at elevated temperatures and pressures prevented use of a fully integrated high-throughput (HTP) screening pipeline. Herein, we report oil the engineering of a novel HTP pretreatment system employing a 96 well-plate format that withstands extreme pretreatment conditions for rapid screening of biomass-enzyme-pretreatment combinations. This includes the development of new approaches to steam heating and water quenching the system that result ill much faster heat Lip and cool clown than previously possible and show consistent temperature histories across the multiwell plate. Coupled pretreatment and enzymatic hydrolysis performance of the well plate pretreatment system is shown to be consistent among the many wells in the device and also with performance of conventional tubular reactors. Biotechnol. Bioeng. 2010;105: 231-238. (C) 2009 Wiley Periodicals, Inc.