Dowe Nancy
National Renewable Energy Laboratory, Golden, CO, USA.
Methods Mol Biol. 2009;581:233-45. doi: 10.1007/978-1-60761-214-8_15.
Cellulase enzyme is a key cost component in the production of fuels and chemicals from lignocellulosic biomass. Cellulolytic ability of the enzyme preparation is often measured by activity assays using model substrates such as filter paper. Using lignocellulosic biomass as the substrate to assess enzyme performance has the potential of being more process relevant. We describe two procedures that use washed pretreated cellulosic material to measure the efficacy of cellulase enzymes. First, a saccharification assay that measures glucose yield as a function of the amount of cellulase used in the process. And second, the simultaneous saccharification and fermentation (SSF) assay measures cellulase performance by the amount of ethanol produced from enzymatic hydrolysis of the cellulosic material. You can use both assays to screen cellulases under a variety of substrate types, loadings, and process conditions.
纤维素酶是从木质纤维素生物质生产燃料和化学品过程中的关键成本组成部分。酶制剂的纤维素分解能力通常通过使用滤纸等模型底物的活性测定来衡量。使用木质纤维素生物质作为底物来评估酶的性能可能与实际生产过程更相关。我们描述了两种使用洗涤过的预处理纤维素材料来测量纤维素酶功效的方法。第一种是糖化测定法，该方法根据过程中使用的纤维素酶量来测量葡萄糖产量。第二种是同步糖化发酵（SSF）测定法，通过纤维素材料酶水解产生的乙醇量来测量纤维素酶的性能。你可以使用这两种测定法在各种底物类型、负载量和工艺条件下筛选纤维素酶。
