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:
-
- 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
- National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
- 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 Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office
- 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. https://doi.org/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. https://doi.org/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 = {Wed Apr 04 00:00:00 EDT 2018},
month = {Wed Apr 04 00:00:00 EDT 2018}
}
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Microbial production of advanced transportation fuels in non-natural hosts
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Engineering for biofuels: exploiting innate microbial capacity or importing biosynthetic potential?
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Paradigm for industrial strain improvement identifies sodium acetate tolerance loci in Zymomonas mobilis and Saccharomyces cerevisiae
journal, May 2010
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- Proceedings of the National Academy of Sciences, Vol. 107, Issue 23
Microbial production of sabinene—a new terpene-based precursor of advanced biofuel
journal, January 2014
- Zhang, Haibo; Liu, Qiang; Cao, Yujin
- Microbial Cell Factories, Vol. 13, Issue 1
Improved ethanol yield and reduced Minimum Ethanol Selling Price (MESP) by modifying low severity dilute acid pretreatment with deacetylation and mechanical refining: 1) Experimental
journal, January 2012
- Chen, Xiaowen; Tao, Ling; Shekiro, Joseph
- Biotechnology for Biofuels, Vol. 5, Issue 1
Engineering xylose metabolism in triacylglycerol-producing Rhodococcus opacus for lignocellulosic fuel production
journal, January 2013
- Kurosawa, Kazuhiko; Wewetzer, Sandra J.; Sinskey, Anthony J.
- Biotechnology for Biofuels, Vol. 6, Issue 1
Bioconversion of lignocellulose: inhibitors and detoxification
journal, January 2013
- Jönsson, Leif J.; Alriksson, Björn; Nilvebrant, Nils-Olof
- Biotechnology for Biofuels, Vol. 6, Issue 1
Insights into acetate toxicity in Zymomonas mobilis8b using different substrates
journal, September 2014
- Yang, Shihui; Franden, Mary Ann; Brown, Steven D.
- Biotechnology for Biofuels, Vol. 7, Issue 1
Transcriptome analysis of Zymomonas mobilis ZM4 reveals mechanisms of tolerance and detoxification of phenolic aldehyde inhibitors from lignocellulose pretreatment
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- Yi, Xia; Gu, Hanqi; Gao, Qiuqiang
- Biotechnology for Biofuels, Vol. 8, Issue 1
Progress and perspective on lignocellulosic hydrolysate inhibitor tolerance improvement in Zymomonas mobilis
journal, February 2018
- Yang, Yongfu; Hu, Mimi; Tang, Ying
- Bioresources and Bioprocessing, Vol. 5, Issue 1
Connecting lignin-degradation pathway with pre-treatment inhibitor sensitivity of Cupriavidus necator
journal, May 2014
- Wang, Wei; Yang, Shihui; Hunsinger, Glendon B.
- Frontiers in Microbiology, Vol. 5
Current knowledge on isobutanol production with Escherichia coli , Bacillus subtilis and Corynebacterium glutamicum
journal, November 2011
- Blombach, Bastian; Eikmanns, Bernhard J.
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Works referencing / citing this record:
Development of Rhodococcus opacus as a chassis for lignin valorization and bioproduction of high-value compounds
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Physiological mechanism of improved tolerance of Saccharomyces cerevisiae to lignin-derived phenolic acids in lignocellulosic ethanol fermentation by short-term adaptation
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Physiological mechanism of improved tolerance of Saccharomyces cerevisiae to lignin-derived phenolic acids in lignocellulosic ethanol fermentation by short-term adaptation
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