High-throughput enzymatic hydrolysis of lignocellulosic biomass via in-situ regeneration

被引：24

作者：

Bharadwaj, Rajiv ^{[1
,3
]}

Wong, April ^{[1
,3
]}

Knierim, Bernhard ^{[1
]}

Singh, Seema ^{[2
,4
]}

Holmes, Bradley M. ^{[2
,4
]}

Auer, Manfred ^{[1
]}

Simmons, Blake A. ^{[2
,4
]}

Adams, Paul D. ^{[1
]}

Singh, Anup K. ^{[1
,3
]}

机构：

[1] Joint BioEnergy Inst, Div Technol, Emeryville, CA 94608 USA

[2] Joint BioEnergy Inst, Deconstruct Div, Emeryville, CA 94608 USA

[3] Sandia Natl Labs, Biosyst Res & Dev Dept, Livermore, CA USA

[4] Sandia Natl Labs, Biomass Sci & Convers Technol Dept, Livermore, CA USA

来源：

BIORESOURCE TECHNOLOGY | 2011年 / 102卷 / 02期

基金：

美国能源部;

关键词：

High-throughput; Cellulase; Ionic liquid; Conductivity; Biofuel; IONIC LIQUID PRETREATMENT; FILTER-PAPER ASSAY; PHOSPHORIC-ACID; CELLULASE; DISSOLUTION; LIGNIN; ADSORPTION; BIOFUELS; KINETICS; DILUTE;

D O I：

10.1016/j.biortech.2010.08.108

中图分类号：

S2 [农业工程];

学科分类号：

082806 [农业信息与电气工程];

摘要：

The high cost of lignocellulolytic enzymes is one of the main barriers towards the development of economically competitive biorefineries. Enzyme engineering can be used to significantly increase the production rate as well as specific activity of enzymes. However, the success of enzyme optimization efforts is currently limited by a lack of robust high-throughput (HIP) cellulase screening platforms for insoluble pretreated lignocellulosic substrates. We have developed a cost-effective microplate based HTP enzyme-screening platform for ionic liquid (IL) pretreated lignocellulose. By performing in-situ biomass regeneration in micro-volumes, we can volumetrically meter biomass (sub-mg loading) and also precisely control the amount of residual IL for engineering novel IL-tolerant cellulases. Our platform only requires straightforward liquid-handling steps and allows the integration of biomass regeneration, washing, saccharification, and imaging steps in a single microtiter plate. The proposed method can be used to screen individual cellulases as well as to develop novel cellulase cocktails. (C) 2010 Elsevier Ltd. All rights reserved.

引用

页码：1329 / 1337

页数：9

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