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预商业化纤维素酶水解预处理玉米秸秆的比较性能。

Comparative performance of precommercial cellulases hydrolyzing pretreated corn stover.

机构信息

National Bioenergy Center, National Renewable Energy Laboratory, Golden, CO, USA.

出版信息

Biotechnol Biofuels. 2011 Sep 7;4:29. doi: 10.1186/1754-6834-4-29.

Abstract

BACKGROUND

Cellulases and related hydrolytic enzymes represent a key cost factor for biochemical conversion of cellulosic biomass feedstocks to sugars for biofuels and chemicals production. The US Department of Energy (DOE) is cost sharing projects to decrease the cost of enzymes for biomass saccharification. The performance of benchmark cellulase preparations produced by Danisco, DSM, Novozymes and Verenium to convert pretreated corn stover (PCS) cellulose to glucose was evaluated under common experimental conditions and is reported here in a non-attributed manner.

RESULTS

Two hydrolysis modes were examined, enzymatic hydrolysis (EH) of PCS whole slurry or washed PCS solids at pH 5 and 50°C, and simultaneous saccharification and fermentation (SSF) of washed PCS solids at pH 5 and 38°C. Enzymes were dosed on a total protein mass basis, with protein quantified using both the bicinchoninic acid (BCA) assay and the Bradford assay. Substantial differences were observed in absolute cellulose to glucose conversion performance levels under the conditions tested. Higher cellulose conversion yields were obtained using washed solids compared to whole slurry, and estimated enzyme protein dosages required to achieve a particular cellulose conversion to glucose yield were extremely dependent on the protein assay used. All four enzyme systems achieved glucose yields of 90% of theoretical or higher in SSF mode. Glucose yields were reduced in EH mode, with all enzymes achieving glucose yields of at least 85% of theoretical on washed PCS solids and 75% in PCS whole slurry. One of the enzyme systems ('enzyme B') exhibited the best overall performance. However in attaining high conversion yields at lower total enzyme protein loadings, the relative and rank ordered performance of the enzyme systems varied significantly depending upon which hydrolysis mode and protein assay were used as the basis for comparison.

CONCLUSIONS

This study provides extensive information about the performance of four precommercial cellulase preparations. Though test conditions were not necessarily optimal for some of the enzymes, all were able to effectively saccharify PCS cellulose. Large differences in the estimated enzyme dosage requirements depending on the assay used to measure protein concentration highlight the need for better consensus methods to quantify enzyme protein.

摘要

背景

纤维素酶和相关水解酶是生物转化纤维素生物质原料为糖生产生物燃料和化学品的关键成本因素。美国能源部(DOE)正在共同承担降低酶制剂用于生物质糖化成本的项目。本文以非归因的方式报告了丹麦诺维信(Danisco)、帝斯曼(DSM)、诺维信(Novozymes)和维美德(Verenium)生产的基准纤维素酶制剂在预处理玉米秸秆(PCS)纤维素转化为葡萄糖方面的性能,这些酶制剂在常见实验条件下进行了评估。

结果

考察了两种水解模式,即 pH5 和 50°C 下的 PCS 全浆或洗涤 PCS 固体的酶水解(EH),以及 pH5 和 38°C 下洗涤 PCS 固体的同步糖化和发酵(SSF)。酶制剂是基于总蛋白质量投加的,蛋白质量使用二辛可宁酸(BCA)法和考马斯亮蓝(Bradford)法进行定量。在测试条件下,观察到绝对纤维素向葡萄糖转化率水平存在显著差异。与全浆相比,洗涤后的固体能获得更高的纤维素转化率,而达到特定纤维素转化率所需的估计酶蛋白剂量则取决于所使用的蛋白分析方法。所有四种酶系统在 SSF 模式下均能达到 90%或更高的理论葡萄糖收率。在 EH 模式下,葡萄糖收率降低,所有酶在洗涤 PCS 固体上的葡萄糖收率至少为理论值的 85%,而在 PCS 全浆中的葡萄糖收率为 75%。有一种酶系统(“酶 B”)表现出最佳的整体性能。然而,在以较低的总酶蛋白负荷获得高转化率时,根据所使用的水解模式和蛋白分析方法作为比较基础,酶系统的相对和排序性能差异显著。

结论

本研究提供了关于四种预商业化纤维素酶制剂性能的广泛信息。虽然一些酶的测试条件不一定是最佳的,但所有酶都能够有效地糖化 PCS 纤维素。由于用于测量蛋白浓度的分析方法不同,估计的酶剂量需求存在很大差异,这突出了需要更好的共识方法来定量酶蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91d5/3200994/6b7eec1c317e/1754-6834-4-29-1.jpg

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