Yang Shihui, Franden Mary Ann, Yang Qing, Chou Yat-Chen, Zhang Min, Pienkos Philip T
Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Environmental Microbial Technology Center of Hubei Province, Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei University, Wuhan, China.
National Bioenergy Center, National Renewable Energy Laboratory, Golden, CO, United States.
Front Bioeng Biotechnol. 2018 Apr 4;6:23. doi: 10.3389/fbioe.2018.00023. eCollection 2018.
The aim of this work was to identify inhibitors in pretreated lignocellulosic slurries, evaluate high-throughput screening strategies, and investigate the impact of inhibitors on potential hydrocarbon-producing microorganisms. Compounds present in slurries that could inhibit microbial growth were identified through a detailed analysis of saccharified slurries by applying a combination of approaches of high-performance liquid chromatography, GC-MS, LC-DAD-MS, and ICP-MS. Several high-throughput assays were then evaluated to generate toxicity profiles. Our results demonstrated that Bioscreen C was useful for analyzing bacterial toxicity but not for yeast. AlamarBlue reduction assay can be a useful high-throughput assay for both bacterial and yeast strains as long as medium components do not interfere with fluorescence measurements. In addition, this work identified two major inhibitors (furfural and ammonium acetate) for three potential hydrocarbon-producing bacterial species that include , and PD630, which are also the primary inhibitors for ethanologens. This study was strived to establish a pipeline to quantify inhibitory compounds in biomass slurries and high-throughput approaches to investigate the effect of inhibitors on microbial biocatalysts, which can be applied for various biomass slurries or hydrolyzates generated through different pretreatment and enzymatic hydrolysis processes or different microbial candidates.
这项工作的目的是鉴定预处理木质纤维素浆料中的抑制剂，评估高通量筛选策略，并研究抑制剂对潜在产烃微生物的影响。通过应用高效液相色谱、气相色谱 - 质谱联用、液相色谱 - 二极管阵列 - 质谱联用和电感耦合等离子体质谱联用等多种方法对糖化浆料进行详细分析，确定了浆料中可能抑制微生物生长的化合物。然后评估了几种高通量测定方法以生成毒性谱。我们的结果表明，Bioscreen C 对于分析细菌毒性有用，但对酵母无用。只要培养基成分不干扰荧光测量，AlamarBlue 还原测定法对细菌和酵母菌株都可以是一种有用的高通量测定法。此外，这项工作确定了三种潜在产烃细菌物种（包括 、 和 PD630）的两种主要抑制剂（糠醛和醋酸铵），它们也是乙醇产生菌 的主要抑制剂。本研究致力于建立一种量化生物质浆料中抑制性化合物的流程以及高通量方法来研究抑制剂对微生物生物催化剂的影响，这可应用于通过不同预处理和酶水解过程产生的各种生物质浆料或水解产物，或不同的微生物候选物。
