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

Yang, Shihui; Franden, Mary A; Yang, Qing; Chou, Yat-Chen; Zhang, Min; Pienkos, Philip T

doi:10.3389/fbioe.2018.00023

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

Journal Article · Wed Apr 04 00:00:00 EDT 2018 · Frontiers in Bioengineering and Biotechnology

DOI:https://doi.org/10.3389/fbioe.2018.00023· OSTI ID:1436074

Yang, Shihui ^[1]; Franden, Mary A ^[2]; Yang, Qing ^[3]; Chou, Yat-Chen ^[2]; Zhang, Min ^[2]; Pienkos, Philip T ^[2]

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

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.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1436074

Report Number(s):: NREL/JA-5100-71459

Journal Information:: Frontiers in Bioengineering and Biotechnology, Vol. 6; ISSN 2296-4185

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 15 works

Citation information provided by
Web of Science

References (52)

Pilot scale pretreatment of wheat straw and comparative evaluation of commercial enzyme preparations for biomass saccharification and fermentation Agrawal, Ruchi; Satlewal, Alok; Gaur, Ruchi Biochemical Engineering Journal, Vol. 102 https://doi.org/10.1016/j.bej.2015.02.018	journal	October 2015
Advanced biofuel production by the yeast Saccharomyces cerevisiae Buijs, Nicolaas A.; Siewers, Verena; Nielsen, Jens Current Opinion in Chemical Biology, Vol. 17, Issue 3 https://doi.org/10.1016/j.cbpa.2013.03.036	journal	June 2013
Pathways for biomass-derived lignin to hydrocarbon fuels Laskar, Dhrubojyoti D.; Yang, Bin; Wang, Huamin Biofuels, Bioproducts and Biorefining, Vol. 7, Issue 5 https://doi.org/10.1002/bbb.1422	journal	July 2013
Carbon source utilization and inhibitor tolerance of 45 oleaginous yeast species Sitepu, Irnayuli; Selby, Tylan; Lin, Ting Journal of Industrial Microbiology & Biotechnology, Vol. 41, Issue 7 https://doi.org/10.1007/s10295-014-1447-y	journal	May 2014
Bioconversion of lignocellulose: inhibitors and detoxification Jönsson, Leif J.; Alriksson, Björn; Nilvebrant, Nils-Olof Biotechnology for Biofuels, Vol. 6, Issue 1 https://doi.org/10.1186/1754-6834-6-16	journal	January 2013
Dilute-Sulfuric Acid Pretreatment of Corn Stover in Pilot-Scale Reactor: Investigation of Yields, Kinetics, and Enzymatic Digestibilities of Solids Schell, Daniel J.; Farmer, Jody; Newman, Millie Applied Biochemistry and Biotechnology, Vol. 105, Issue 1-3, p. 69-86 https://doi.org/10.1385/ABAB:105:1-3:69	journal	January 2003
Transcriptome analysis of Zymomonas mobilis ZM4 reveals mechanisms of tolerance and detoxification of phenolic aldehyde inhibitors from lignocellulose pretreatment Yi, Xia; Gu, Hanqi; Gao, Qiuqiang Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0333-9	journal	September 2015
Perspective and Prospective of Pretreatment of Corn Straw for Butanol Production Baral, Nawa Raj; Li, Jiangzheng; Jha, Ajay Kumar Applied Biochemistry and Biotechnology, Vol. 172, Issue 2 https://doi.org/10.1007/s12010-013-0548-9	journal	October 2013
Exploration of Natural Biomass Utilization Systems (NBUS) for advanced biofuel—from systems biology to synthetic design Xie, Shangxian; Syrenne, Ryan; Sun, Su Current Opinion in Biotechnology, Vol. 27 https://doi.org/10.1016/j.copbio.2014.02.007	journal	June 2014
Pretreatment of lignocellulose: Formation of inhibitory by-products and strategies for minimizing their effects Jönsson, Leif J.; Martín, Carlos Bioresource Technology, Vol. 199 https://doi.org/10.1016/j.biortech.2015.10.009	journal	January 2016
Poly(3-hydroxybutyrate) production by Cupriavidus necator using waste glycerol Cavalheiro, João M. B. T.; de Almeida, M. Catarina M. D.; Grandfils, Christian Process Biochemistry, Vol. 44, Issue 5 https://doi.org/10.1016/j.procbio.2009.01.008	journal	May 2009
Efficient coagulation of microalgae in cultures with filamentous fungi Xie, Shangxian; Sun, Su; Dai, Susie Y. Algal Research, Vol. 2, Issue 1 https://doi.org/10.1016/j.algal.2012.11.004	journal	January 2013
Microbial production of advanced transportation fuels in non-natural hosts Connor, Michael R.; Liao, James C. Current Opinion in Biotechnology, Vol. 20, Issue 3, p. 307-315 https://doi.org/10.1016/j.copbio.2009.04.002	journal	June 2009
Inhibition of growth of Zymomonas mobilis by model compounds found in lignocellulosic hydrolysates Franden, Mary Ann; Pilath, Heidi M.; Mohagheghi, Ali Biotechnology for Biofuels, Vol. 6, Issue 1 https://doi.org/10.1186/1754-6834-6-99	journal	January 2013
Identification and microbial production of a terpene-based advanced biofuel Peralta-Yahya, Pamela P.; Ouellet, Mario; Chan, Rossana Nature Communications, Vol. 2, Issue 1 https://doi.org/10.1038/ncomms1494	journal	September 2011
Rhodococcus opacus B4: a promising bacterium for production of biofuels and biobased chemicals Castro, Ana Rita; Rocha, Isabel; Alves, Maria Madalena AMB Express, Vol. 6, Issue 1 https://doi.org/10.1186/s13568-016-0207-y	journal	May 2016
Engineering Corynebacterium glutamicum for isobutanol production Smith, Kevin Michael; Cho, Kwang-Myung; Liao, James C. Applied Microbiology and Biotechnology, Vol. 87, Issue 3 https://doi.org/10.1007/s00253-010-2522-6	journal	April 2010
Engineering Terpene Biosynthesis in Streptomyces for Production of the Advanced Biofuel Precursor Bisabolene Phelan, Ryan M.; Sekurova, Olga N.; Keasling, Jay D. ACS Synthetic Biology, Vol. 4, Issue 4 https://doi.org/10.1021/sb5002517	journal	July 2014
High-cell-density batch fermentation of Rhodococcus opacus PD630 using a high glucose concentration for triacylglycerol production Kurosawa, Kazuhiko; Boccazzi, Paolo; de Almeida, Naomi M. Journal of Biotechnology, Vol. 147, Issue 3-4 https://doi.org/10.1016/j.jbiotec.2010.04.003	journal	June 2010
Lipid and fatty acid metabolism in Ralstonia eutropha: relevance for the biotechnological production of value-added products Riedel, Sebastian L.; Lu, Jingnan; Stahl, Ulf Applied Microbiology and Biotechnology, Vol. 98, Issue 4 https://doi.org/10.1007/s00253-013-5430-8	journal	December 2013
Improved saccharification of pilot-scale acid pretreated wheat straw by exploiting the synergistic behavior of lignocellulose degrading enzymes Agrawal, Ruchi; Gaur, Ruchi; Mathur, Anshu RSC Advances, Vol. 5, Issue 87 https://doi.org/10.1039/C5RA13360B	journal	January 2015
The role of pretreatment in improving the enzymatic hydrolysis of lignocellulosic materials Sun, Shaoni; Sun, Shaolong; Cao, Xuefei Bioresource Technology, Vol. 199 https://doi.org/10.1016/j.biortech.2015.08.061	journal	January 2016
Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels Atsumi, Shota; Hanai, Taizo; Liao, James C. Nature, Vol. 451, Issue 7174, p. 86-89 https://doi.org/10.1038/nature06450	journal	January 2008
Bacteria engineered for fuel ethanol production: current status Dien, B. S.; Cotta, M. A.; Jeffries, T. W. Applied Microbiology and Biotechnology, Vol. 63, Issue 3, p. 258-266 https://doi.org/10.1007/s00253-003-1444-y	journal	December 2003
Paradigm for industrial strain improvement identifies sodium acetate tolerance loci in Zymomonas mobilis and Saccharomyces cerevisiae Yang, S.; Land, M. L.; Klingeman, D. M. Proceedings of the National Academy of Sciences, Vol. 107, Issue 23 https://doi.org/10.1073/pnas.0914506107	journal	May 2010
Investigating the enzyme-lignin binding with surfactants for improved saccharification of pilot scale pretreated wheat straw Agrawal, Ruchi; Satlewal, Alok; Kapoor, Manali Bioresource Technology, Vol. 224 https://doi.org/10.1016/j.biortech.2016.11.026	journal	January 2017
Lipid Production from Dilute Alkali Corn Stover Lignin by Rhodococcus Strains He, Yucai; Li, Xiaolu; Ben, Haoxi ACS Sustainable Chemistry & Engineering, Vol. 5, Issue 3 https://doi.org/10.1021/acssuschemeng.6b02627	journal	January 2017
Improving furfural tolerance of Zymomonas mobilis by rewiring a sigma factor RpoD protein Tan, Fu-Rong; Dai, Li-Chun; Wu, Bo Applied Microbiology and Biotechnology, Vol. 99, Issue 12 https://doi.org/10.1007/s00253-015-6577-2	journal	April 2015
Current knowledge on isobutanol production with Escherichia coli , Bacillus subtilis and Corynebacterium glutamicum Blombach, Bastian; Eikmanns, Bernhard J. Bioengineered Bugs, Vol. 2, Issue 6 https://doi.org/10.4161/bbug.2.6.17845	journal	November 2011
Insights into acetate toxicity in Zymomonas mobilis8b using different substrates Yang, Shihui; Franden, Mary Ann; Brown, Steven D. Biotechnology for Biofuels, Vol. 7, Issue 1 https://doi.org/10.1186/s13068-014-0140-8	journal	September 2014
Features of promising technologies for pretreatment of lignocellulosic biomass Mosier, Nathan; Wyman, Charles; Dale, Bruce Bioresource Technology, Vol. 96, Issue 6, p. 673-686 https://doi.org/10.1016/j.biortech.2004.06.025	journal	April 2005
Methods for Pretreatment of Lignocellulosic Biomass for Efficient Hydrolysis and Biofuel Production Kumar, Parveen; Barrett, Diane M.; Delwiche, Michael J. Industrial & Engineering Chemistry Research, Vol. 48, Issue 8, p. 3713-3729 https://doi.org/10.1021/ie801542g	journal	April 2009
Integrated approach for effective bioethanol production using whole slurry from autohydrolyzed Eucalyptus globulus wood at high-solid loadings Romaní, Aloia; Ruiz, Héctor A.; Pereira, Francisco B. Fuel, Vol. 135 https://doi.org/10.1016/j.fuel.2014.06.061	journal	November 2014
Bio-ethanol – the fuel of tomorrow from the residues of today Hahn-Hägerdal, B.; Galbe, M.; Gorwa-Grauslund, M. F. Trends in Biotechnology, Vol. 24, Issue 12, p. 549-556 https://doi.org/10.1016/j.tibtech.2006.10.004	journal	December 2006
Improved ethanol yield and reduced Minimum Ethanol Selling Price (MESP) by modifying low severity dilute acid pretreatment with deacetylation and mechanical refining: 1) Experimental Chen, Xiaowen; Tao, Ling; Shekiro, Joseph Biotechnology for Biofuels, Vol. 5, Issue 1 https://doi.org/10.1186/1754-6834-5-60	journal	January 2012
DMR (deacetylation and mechanical refining) processing of corn stover achieves high monomeric sugar concentrations (230 g L ⁻¹ ) during enzymatic hydrolysis and high ethanol concentrations (>10% v/v) during fermentation without hydrolysate purification or concentration Chen, Xiaowen; Kuhn, Erik; Jennings, Edward W. Energy & Environmental Science, Vol. 9, Issue 4 https://doi.org/10.1039/C5EE03718B	journal	January 2016
Green methods of lignocellulose pretreatment for biorefinery development Capolupo, Laura; Faraco, Vincenza Applied Microbiology and Biotechnology, Vol. 100, Issue 22 https://doi.org/10.1007/s00253-016-7884-y	journal	October 2016
Synergistic enzymatic and microbial lignin conversion Zhao, Cheng; Xie, Shangxian; Pu, Yunqiao Green Chemistry, Vol. 18, Issue 5 https://doi.org/10.1039/C5GC01955A	journal	January 2016
The Zymomonas mobilis regulator hfq contributes to tolerance against multiple lignocellulosic pretreatment inhibitors Yang, Shihui; Pelletier, Dale A.; Lu, Tse-Yuan S. BMC Microbiology, Vol. 10, Article No. 135 https://doi.org/10.1186/1471-2180-10-135	journal	January 2010
Engineering for biofuels: exploiting innate microbial capacity or importing biosynthetic potential? Alper, Hal; Stephanopoulos, Gregory Nature Reviews Microbiology, Vol. 7, Issue 10 https://doi.org/10.1038/nrmicro2186	journal	October 2009
Bioconversion of lignin model compounds with oleaginous Rhodococci Kosa, Matyas; Ragauskas, Arthur J. Applied Microbiology and Biotechnology, Vol. 93, Issue 2 https://doi.org/10.1007/s00253-011-3743-z	journal	December 2011
Development of a high-throughput method to evaluate the impact of inhibitory compounds from lignocellulosic hydrolysates on the growth of Zymomonas mobilis Franden, Mary Ann; Pienkos, Philip T.; Zhang, Min Journal of Biotechnology, Vol. 144, Issue 4 https://doi.org/10.1016/j.jbiotec.2009.08.006	journal	December 2009
Microbial lipid production by oleaginous Rhodococci cultured in lignocellulosic autohydrolysates Wei, Zhen; Zeng, Guangming; Huang, Fang Applied Microbiology and Biotechnology, Vol. 99, Issue 17 https://doi.org/10.1007/s00253-015-6752-5	journal	July 2015
Modeling and optimization of the dilute-sulfuric-acid pretreatment of corn stover, poplar and switchgrass Esteghlalian, Alireza; Hashimoto, Andrew G.; Fenske, John J. Bioresource Technology, Vol. 59, Issue 2-3, p. 129-136 https://doi.org/10.1016/S0960-8524(97)81606-9	journal	February 1997
Engineering xylose metabolism in triacylglycerol-producing Rhodococcus opacus for lignocellulosic fuel production Kurosawa, Kazuhiko; Wewetzer, Sandra J.; Sinskey, Anthony J. Biotechnology for Biofuels, Vol. 6, Issue 1 https://doi.org/10.1186/1754-6834-6-134	journal	January 2013
Connecting lignin-degradation pathway with pre-treatment inhibitor sensitivity of Cupriavidus necator Wang, Wei; Yang, Shihui; Hunsinger, Glendon B. Frontiers in Microbiology, Vol. 5 https://doi.org/10.3389/fmicb.2014.00247	journal	May 2014
Advanced biofuel production in microbes Peralta-Yahya, Pamela P.; Keasling, Jay D. Biotechnology Journal, Vol. 5, Issue 2, p. 147-162 https://doi.org/10.1002/biot.200900220	journal	January 2010
Microbial production of sabinene—a new terpene-based precursor of advanced biofuel Zhang, Haibo; Liu, Qiang; Cao, Yujin Microbial Cell Factories, Vol. 13, Issue 1 https://doi.org/10.1186/1475-2859-13-20	journal	January 2014
Effect of hemicellulose liquid phase on the enzymatic hydrolysis of autohydrolyzed Eucalyptus globulus wood Romaní, Aloia; Ruiz, Héctor A.; Pereira, Francisco B. Biomass Conversion and Biorefinery, Vol. 4, Issue 2 https://doi.org/10.1007/s13399-013-0093-3	journal	August 2013
Current Pretreatment Technologies for the Development of Cellulosic Ethanol and Biorefineries Silveira, Marcos Henrique Luciano; Morais, Ana Rita C.; da Costa Lopes, Andre M. ChemSusChem, Vol. 8, Issue 20 https://doi.org/10.1002/cssc.201500282	journal	September 2015
Progress and perspective on lignocellulosic hydrolysate inhibitor tolerance improvement in Zymomonas mobilis Yang, Yongfu; Hu, Mimi; Tang, Ying Bioresources and Bioprocessing, Vol. 5, Issue 1 https://doi.org/10.1186/s40643-018-0193-9	journal	February 2018
Performance of a newly developed integrant of Zymomonas mobilis for ethanol production on corn stover hydrolysate Mohagheghi, Ali; Dowe, Nancy; Schell, Daniel Biotechnology Letters, Vol. 26, Issue 4 https://doi.org/10.1023/b:bile.0000015451.96737.96	journal	February 2004

Cited By (2)

Development of Rhodococcus opacus as a chassis for lignin valorization and bioproduction of high-value compounds Anthony, Winston E.; Carr, Rhiannon R.; DeLorenzo, Drew M. Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1535-3	journal	August 2019
Physiological mechanism of improved tolerance of Saccharomyces cerevisiae to lignin-derived phenolic acids in lignocellulosic ethanol fermentation by short-term adaptation Gu, Hanqi; Zhu, Yuyong; Peng, Yanfang Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1610-9	journal	November 2019

Similar Records

Connecting Lignin-Degradation Pathway with Pre-Treatment Inhibitor Sensitivity of Cupriavidus necator

Journal Article · Tue May 27 00:00:00 EDT 2014 · Frontiers in Microbiology · OSTI ID:1436074

Wang, W.; Yang, S.; Hunsinger, G. B.; +2 more

Biological Production of a Hydrocarbon Fuel Intermediate Polyhydroxybutyrate (Phb) from a Process Relevant Lignocellulosic Derived Sugar

Conference · Sun Mar 22 00:00:00 EDT 2015 · OSTI ID:1436074

Wang, Wei; Mohagheghi, Ali; Mittal, Ashutosh; +2 more

Biological Production of a Hydrocarbon Fuel Intermediate Polyhydroxybutyrate (PHB) from a Process Relevant Lignocellulosic Derived Sugar (Poster)

Conference · Tue Apr 01 00:00:00 EDT 2014 · OSTI ID:1436074

Wang, W.; Mittal, A.; Mohagheghi, A.; +1 more

Related Subjects

09 BIOMASS FUELS
saccharified slurry
inhibitor
furfural
acetate
high-throughput screening

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

Citation Formats

References (52)

Cited By (2)

Similar Records

Related Subjects