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Title: Identification of Inhibitors in Lignocellulosic Slurries and Determination of Their Effect on Hydrocarbon-Producing Microorganisms

Abstract

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 Escherichia coli, Cupriavidus necator, and Rhodococcus opacus PD630, which are also the primary inhibitors for ethanologens. Here, 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.

Authors:
 [1];  [2];  [3];  [2];  [2];  [2]
  1. Hubei Univ., Wuhan (China). College of Life Sciences, Hubei Key Lab. of Industrial Biotechnology, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Environmental Microbial Technology Center of Hubei Province; National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
  3. Hubei Univ., Wuhan (China). College of Life Sciences, Hubei Key Lab. of Industrial Biotechnology, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Environmental Microbial Technology Center of Hubei Province
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B)
OSTI Identifier:
1436074
Report Number(s):
NREL/JA-5100-71459
Journal ID: ISSN 2296-4185
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Bioengineering and Biotechnology
Additional Journal Information:
Journal Volume: 6; Journal ID: ISSN 2296-4185
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; saccharified slurry; inhibitor; furfural; acetate; high-throughput screening

Citation Formats

Yang, Shihui, Franden, Mary A, Yang, Qing, Chou, Yat-Chen, Zhang, Min, and Pienkos, Philip T. Identification of Inhibitors in Lignocellulosic Slurries and Determination of Their Effect on Hydrocarbon-Producing Microorganisms. United States: N. p., 2018. Web. doi:10.3389/fbioe.2018.00023.
Yang, Shihui, Franden, Mary A, Yang, Qing, Chou, Yat-Chen, Zhang, Min, & Pienkos, Philip T. Identification of Inhibitors in Lignocellulosic Slurries and Determination of Their Effect on Hydrocarbon-Producing Microorganisms. United States. doi:10.3389/fbioe.2018.00023.
Yang, Shihui, Franden, Mary A, Yang, Qing, Chou, Yat-Chen, Zhang, Min, and Pienkos, Philip T. Wed . "Identification of Inhibitors in Lignocellulosic Slurries and Determination of Their Effect on Hydrocarbon-Producing Microorganisms". United States. doi:10.3389/fbioe.2018.00023. https://www.osti.gov/servlets/purl/1436074.
@article{osti_1436074,
title = {Identification of Inhibitors in Lignocellulosic Slurries and Determination of Their Effect on Hydrocarbon-Producing Microorganisms},
author = {Yang, Shihui and Franden, Mary A and Yang, Qing and Chou, Yat-Chen and Zhang, Min and Pienkos, Philip T},
abstractNote = {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 Escherichia coli, Cupriavidus necator, and Rhodococcus opacus PD630, which are also the primary inhibitors for ethanologens. Here, 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.},
doi = {10.3389/fbioe.2018.00023},
journal = {Frontiers in Bioengineering and Biotechnology},
number = ,
volume = 6,
place = {United States},
year = {2018},
month = {4}
}

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