Methodology for enabling high-throughput simultaneous saccharification and fermentation screening of yeast using solid biomass as a substrate.
Methodology for enabling high-throughput simultaneous saccharification and fermentation screening of yeast using solid biomass as a substrate.
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DOI：
10.1186/s13068-014-0181-z
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发表时间：
2015
影响因子：
6.3
通讯作者：
Waldron KW
中科院分区：
文献类型：
--
作者：
Elliston A;Wood IP;Soucouri MJ;Tantale RJ;Dicks J;Roberts IN;Waldron KW
高通量(HTP)筛查正在成为整理生物数据的日益有用的工具，否则将需要使用过多的资源。第二代生物燃料生产就是这样一个过程。HTP筛查允许以更快的速度和成本效益进行大样本集的调查。本文概述了一种方法，它将使固体木质纤维底物在96孔板规模上进行水解和发酵，促进乙醇生产的HTP筛选，同时保持与更大规模实现的重复性相似的重复性。结果表明，使用密度一致的生物质薄片(传单)作为纸张，以及可以用管道输送的经过处理的生物质浆液，可以实现标准化和准确地转移到96孔板(分别为±3.1%和1.7%)。用不同体积的同时糖化发酵(SSF)处理这些底物，无论是在标准摇瓶(200m L)、通用瓶(10m L)和96孔板(1m L)上，最终的乙醇产量都没有显著差异。底物浓度高达10%(w/v)的SSFS在1mL体积下成功地进行了试验。通过对油菜秸秆和小麦秸秆进行蒸汽爆破处理，验证了该方法的有效性。这种方法可以用相对快速的96孔板SSF分析代替大的摇瓶反应，从而允许HTP实验。此外，该方法与许多标准化检测技术兼容，例如简单比色法、高效液相色谱(HPLC)和核磁共振(NMR)光谱。此外，这项研究通过允许HTP SSF筛选，在第二代生物燃料和新型生物基础化学品的生物质基质的生物精炼中具有实际应用，这将使选定的样品能够扩大规模或进行更详细的研究。
High-throughput (HTP) screening is becoming an increasingly useful tool for collating biological data which would otherwise require the employment of excessive resources. Second generation biofuel production is one such process. HTP screening allows the investigation of large sample sets to be undertaken with increased speed and cost effectiveness. This paper outlines a methodology that will enable solid lignocellulosic substrates to be hydrolyzed and fermented at a 96-well plate scale, facilitating HTP screening of ethanol production, whilst maintaining repeatability similar to that achieved at a larger scale. The results showed that utilizing sheets of biomass of consistent density (handbills), for paper, and slurries of pretreated biomass that could be pipetted allowed standardized and accurate transfers to 96-well plates to be achieved (±3.1 and 1.7%, respectively). Processing these substrates by simultaneous saccharification and fermentation (SSF) at various volumes showed no significant difference on final ethanol yields, either at standard shake flask (200 mL), universal bottle (10 mL) or 96-well plate (1 mL) scales. Substrate concentrations of up to 10% (w/v) were trialed successfully for SSFs at 1 mL volume. The methodology was successfully tested by showing the effects of steam explosion pretreatment on both oilseed rape and wheat straws. This methodology could be used to replace large shake flask reactions with comparatively fast 96-well plate SSF assays allowing for HTP experimentation. Additionally this method is compatible with a number of standardized assay techniques such as simple colorimetric, High-performance liquid chromatography (HPLC) and Nuclear magnetic resonance (NMR) spectroscopy. Furthermore this research has practical uses in the biorefining of biomass substrates for second generation biofuels and novel biobased chemicals by allowing HTP SSF screening, which should allow selected samples to be scaled up or studied in more detail.
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影响因子：
5
作者：
BAILEY, MJ
通讯作者：
BAILEY, MJ
影响因子：
3.6
作者：
Santoro, Nicholas;Cantu, Shane L.;Walton, Jonathan D.
通讯作者：
Walton, Jonathan D.
影响因子：
6.4
作者：
Navarro D;Couturier M;da Silva GG;Berrin JG;Rouau X;Asther M;Bignon C
通讯作者：
Bignon C
影响因子：
3.1
作者：
BLAKENEY, AB;HARRIS, PJ;STONE, BA
通讯作者：
STONE, BA
影响因子：
3.4
作者：
Luo, Xiaolin;Zhu, J. Y.
通讯作者：
Zhu, J. Y.