skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Adaptation to low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and growth of Saccharomyces cerevisiae

Abstract

Here, lignocellulosic bioethanol from renewable feedstocks using Saccharomyces cerevisiae is a promising alternative to fossil fuels owing to environmental challenges. S. cerevisiae is frequently challenged by bacterial contamination and a combination of lignocellulosic inhibitors formed during the pre-treatment, in terms of growth, ethanol yield and productivity. We investigated the phenotypic robustness of a brewing yeast strain TMB3500 and its ability to adapt to low pH thereby preventing bacterial contamination along with lignocellulosic inhibitors by short-term adaptation and adaptive lab evolution (ALE). The short-term adaptation strategy was used to investigate the inherent ability of strain TMB3500 to activate a robust phenotype involving pre-culturing yeast cells in defined medium with lignocellulosic inhibitors at pH 5.0 until late exponential phase prior to inoculating them in defined media with the same inhibitor cocktail at pH 3.7. Adapted cells were able to grow aerobically, ferment anaerobically (glucose exhaustion by 19 +/- 5 h to yield 0.45 +/- 0.01 g ethanol g glucose-1) and portray significant detoxification of inhibitors at pH 3.7, when compared to non-adapted cells. ALE was performed to investigate whether a stable strain could be developed to grow and ferment at low pH with lignocellulosic inhibitors in a continuous suspension culture. Though amore » robust population was obtained after 3600 h with an ability to grow and ferment at pH 3.7 with inhibitors, inhibitor robustness was not stable as indicated by the characterisation of the evolved culture possibly due to phenotypic plasticity. With further research, this short-term adaptation and low pH strategy could be successfully applied in lignocellulosic ethanol plants to prevent bacterial contamination.« less

Authors:
 [1];  [2];  [1];  [1]
  1. Lund Univ., Lund (Sweden)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
Swedish National Energy Agency; USDOE
OSTI Identifier:
1325063
Report Number(s):
NREL/JA-5100-67105
Journal ID: ISSN 2191-0855
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
AMB Express
Additional Journal Information:
Journal Volume: 6; Journal Issue: 1; Journal ID: ISSN 2191-0855
Publisher:
Springer
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; Saccharomyces cerevisiae; low pH; lignocellulosic inhibitors; phenotypic robustness; adaptation; ethanol yield

Citation Formats

Narayanan, Venkatachalam, Sànchez i Nogué, Violeta, van Niel, Ed W. J., and Gorwa-Grauslund, Marie F. Adaptation to low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and growth of Saccharomyces cerevisiae. United States: N. p., 2016. Web. doi:10.1186/s13568-016-0234-8.
Narayanan, Venkatachalam, Sànchez i Nogué, Violeta, van Niel, Ed W. J., & Gorwa-Grauslund, Marie F. Adaptation to low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and growth of Saccharomyces cerevisiae. United States. doi:10.1186/s13568-016-0234-8.
Narayanan, Venkatachalam, Sànchez i Nogué, Violeta, van Niel, Ed W. J., and Gorwa-Grauslund, Marie F. Fri . "Adaptation to low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and growth of Saccharomyces cerevisiae". United States. doi:10.1186/s13568-016-0234-8. https://www.osti.gov/servlets/purl/1325063.
@article{osti_1325063,
title = {Adaptation to low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and growth of Saccharomyces cerevisiae},
author = {Narayanan, Venkatachalam and Sànchez i Nogué, Violeta and van Niel, Ed W. J. and Gorwa-Grauslund, Marie F.},
abstractNote = {Here, lignocellulosic bioethanol from renewable feedstocks using Saccharomyces cerevisiae is a promising alternative to fossil fuels owing to environmental challenges. S. cerevisiae is frequently challenged by bacterial contamination and a combination of lignocellulosic inhibitors formed during the pre-treatment, in terms of growth, ethanol yield and productivity. We investigated the phenotypic robustness of a brewing yeast strain TMB3500 and its ability to adapt to low pH thereby preventing bacterial contamination along with lignocellulosic inhibitors by short-term adaptation and adaptive lab evolution (ALE). The short-term adaptation strategy was used to investigate the inherent ability of strain TMB3500 to activate a robust phenotype involving pre-culturing yeast cells in defined medium with lignocellulosic inhibitors at pH 5.0 until late exponential phase prior to inoculating them in defined media with the same inhibitor cocktail at pH 3.7. Adapted cells were able to grow aerobically, ferment anaerobically (glucose exhaustion by 19 +/- 5 h to yield 0.45 +/- 0.01 g ethanol g glucose-1) and portray significant detoxification of inhibitors at pH 3.7, when compared to non-adapted cells. ALE was performed to investigate whether a stable strain could be developed to grow and ferment at low pH with lignocellulosic inhibitors in a continuous suspension culture. Though a robust population was obtained after 3600 h with an ability to grow and ferment at pH 3.7 with inhibitors, inhibitor robustness was not stable as indicated by the characterisation of the evolved culture possibly due to phenotypic plasticity. With further research, this short-term adaptation and low pH strategy could be successfully applied in lignocellulosic ethanol plants to prevent bacterial contamination.},
doi = {10.1186/s13568-016-0234-8},
journal = {AMB Express},
number = 1,
volume = 6,
place = {United States},
year = {2016},
month = {8}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 3 works
Citation information provided by
Web of Science

Figures / Tables:

Fig. 1 Fig. 1: Effect of low pH with 6 g L−1 acetic acid and individual inhibitors (1.5 g L−1 furfural or 0.5 g L−1 HMF or 1 g L−1 vanillin) on short‑term adapted (black) and non‑adapted (white) cells. pH 3.7 with inhibitors includes the IC

Save / Share:

Works referenced in this record:

Physiology of the fuel ethanol strain Saccharomyces cerevisiae PE-2 at low pH indicates a context-dependent performance relevant for industrial applications
journal, October 2014

  • Della-Bianca, Bianca E.; de Hulster, Erik; Pronk, Jack T.
  • FEMS Yeast Research, Vol. 14, Issue 8
  • DOI: 10.1111/1567-1364.12217

Increased tolerance and conversion of inhibitors in lignocellulosic hydrolysates by Saccharomyces cerevisiae
journal, January 2007

  • Almeida, Joao; Modig, Tobias; Petersson, Anneli
  • Journal of Chemical Technology & Biotechnology, Vol. 82, Issue 4, p. 340-349
  • DOI: 10.1002/jctb.1676

Bioconversion of lignocellulose: inhibitors and detoxification
journal, January 2013

  • Jönsson, Leif J.; Alriksson, Björn; Nilvebrant, Nils-Olof
  • Biotechnology for Biofuels, Vol. 6, Issue 1
  • DOI: 10.1186/1754-6834-6-16

Microbial cell individuality and the underlying sources of heterogeneity
journal, August 2006


Inhibition of ethanol-producing yeast and bacteria by degradation products produced during pre-treatment of biomass
journal, August 2004

  • Klinke, H. B.; Thomsen, A. B.; Ahring, B. K.
  • Applied Microbiology and Biotechnology, Vol. 66, Issue 1, p. 10-26
  • DOI: 10.1007/s00253-004-1642-2

Biofilm production by Zymomonas mobilis enhances ethanol production and tolerance to toxic inhibitors from rice bran hydrolysate
journal, September 2014

  • Todhanakasem, Tatsaporn; Sangsutthiseree, Atit; Areerat, Kamonchanok
  • New Biotechnology, Vol. 31, Issue 5
  • DOI: 10.1016/j.nbt.2014.06.002

Ethanol fermentation of various pretreated and hydrolyzed substrates at low initial pH
journal, April 2007

  • Kádár, Zsófia; Maltha, San Feng; Szengyel, Zsolt
  • Applied Biochemistry and Biotechnology, Vol. 137-140, Issue 1-12
  • DOI: 10.1007/s12010-007-9102-y

Differentiation of Brewery Yeast Strains by dna Fingerprinting
journal, March 1995


Metabolic variability in bioprocessing: implications of microbial phenotypic heterogeneity
journal, December 2014


Identification of furfural as a key toxin in lignocellulosic hydrolysates and evolution of a tolerant yeast strain
journal, November 2008


Microbial contamination of fuel ethanol fermentations: Bioethanol contamination
journal, August 2011


Yeast responses to stresses associated with industrial brewery handling: Figure 1
journal, September 2007


Main and interaction effects of acetic acid, furfural, andp-hydroxybenzoic acid on growth and ethanol productivity of yeasts
journal, April 1999


Adaptation of Microorganisms to Cold Temperatures, Weak Acid Preservatives, Low pH, and Osmotic Stress: A Review
journal, January 2004


Effect of High Solids Loading on Bacterial Contamination in Lignocellulosic Ethanol Production
journal, May 2013


Antimicrobial strategies for limiting bacterial contaminants in fuel bioethanol fermentations
journal, June 2011

  • Muthaiyan, Arunachalam; Limayem, Alya; Ricke, Steven C.
  • Progress in Energy and Combustion Science, Vol. 37, Issue 3
  • DOI: 10.1016/j.pecs.2010.06.005

Flocculation, adhesion and biofilm formation in yeasts
journal, April 2006


Bet Hedging in Yeast by Heterogeneous, Age-Correlated Expression of a Stress Protectant
journal, May 2012


Increasing Anaerobic Acetate Consumption and Ethanol Yields in Saccharomyces cerevisiae with NADPH-Specific Alcohol Dehydrogenase
journal, September 2015

  • Henningsen, Brooks M.; Hon, Shuen; Covalla, Sean F.
  • Applied and Environmental Microbiology, Vol. 81, Issue 23
  • DOI: 10.1128/AEM.01689-15

Yeast population dynamics of industrial fuel-ethanol fermentation process assessed by PCR-fingerprinting
journal, July 2005

  • Silva-Filho, Eurípedes Alves da; Santos, Scheila Karina Brito dos; Resende, Alecsandra do Monte
  • Antonie van Leeuwenhoek, Vol. 88, Issue 1
  • DOI: 10.1007/s10482-004-7283-8

Effect of benzoic acid on metabolic fluxes in yeasts: A continuous-culture study on the regulation of respiration and alcoholic fermentation
journal, July 1992

  • Verduyn, Cornelis; Postma, Erik; Scheffers, W. Alexander
  • Yeast, Vol. 8, Issue 7
  • DOI: 10.1002/yea.320080703

Molecular mechanisms of yeast tolerance and in situ detoxification of lignocellulose hydrolysates
journal, March 2011


Stress-related challenges in pentose fermentation to ethanol by the yeast Saccharomyces cerevisiae
journal, February 2011

  • Almeida, João R. M.; Runquist, David; Sànchez Nogué, Violeta
  • Biotechnology Journal, Vol. 6, Issue 3
  • DOI: 10.1002/biot.201000301

DNA typing methods for differentiation of yeasts related to dry-cured meat products
journal, March 2006


Selective suppression of bacterial contaminants by process conditions during lignocellulose based yeast fermentations
journal, January 2011

  • Albers, Eva; Johansson, Emma; Franzén, Carl Johan
  • Biotechnology for Biofuels, Vol. 4, Issue 1
  • DOI: 10.1186/1754-6834-4-59

Metabolic effects of furaldehydes and impacts on biotechnological processes
journal, January 2009

  • Almeida, João R. M.; Bertilsson, Magnus; Gorwa-Grauslund, Marie F.
  • Applied Microbiology and Biotechnology, Vol. 82, Issue 4
  • DOI: 10.1007/s00253-009-1875-1

The Cytosolic pH of Individual Saccharomyces cerevisiae Cells Is a Key Factor in Acetic Acid Tolerance
journal, September 2015

  • Fernández-Niño, Miguel; Marquina, Maribel; Swinnen, Steve
  • Applied and Environmental Microbiology, Vol. 81, Issue 22
  • DOI: 10.1128/AEM.02313-15

Evolutionary engineering of Saccharomyces cerevisiae for enhanced tolerance to hydrolysates of lignocellulosic biomass : Adaptive Evolution for Hydrolysates Tolerance in Yeast
journal, July 2013

  • Almario, María P.; Reyes, Luis H.; Kao, Katy C.
  • Biotechnology and Bioengineering, Vol. 110, Issue 10
  • DOI: 10.1002/bit.24938

Enhancement of Acetic Acid Tolerance in Saccharomyces cerevisiae by Overexpression of the HAA1 Gene, Encoding a Transcriptional Activator
journal, September 2012

  • Tanaka, Koichi; Ishii, Yukari; Ogawa, Jun
  • Applied and Environmental Microbiology, Vol. 78, Issue 22
  • DOI: 10.1128/AEM.02356-12

Short-term adaptation during propagation improves the performance of xylose-fermenting Saccharomyces cerevisiae in simultaneous saccharification and co-fermentation
journal, December 2015

  • Nielsen, Fredrik; Tomás-Pejó, Elia; Olsson, Lisbeth
  • Biotechnology for Biofuels, Vol. 8, Issue 1
  • DOI: 10.1186/s13068-015-0399-4

Editorial overview: Energy biotechnology
journal, June 2015


Variability of the response ofSaccharomyces cerevisiae strains to lignocellulose hydrolysate
journal, January 2008

  • Modig, Tobias; Almeida, João R. M.; Gorwa-Grauslund, Marie F.
  • Biotechnology and Bioengineering, Vol. 100, Issue 3
  • DOI: 10.1002/bit.21789

Biotechnological strategies to overcome inhibitors in lignocellulose hydrolysates for ethanol production: review
journal, May 2010


Low external pH induces HOG1-dependent changes in the organization of the Saccharomyces cerevisiae cell wall
journal, January 2001


Reduction of invasive bacteria in ethanol fermentations using bacteriophages: Reduction of Invasive Bacteria Using Phages
journal, June 2015

  • Worley-Morse, Thomas O.; Deshusses, Marc A.; Gunsch, Claudia K.
  • Biotechnology and Bioengineering, Vol. 112, Issue 8
  • DOI: 10.1002/bit.25586

Overexpression of Saccharomyces cerevisiae transcription factor and multidrug resistance genes conveys enhanced resistance to lignocellulose-derived fermentation inhibitors
journal, February 2010


DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting
journal, January 1992

  • Akopyanz, Natalia; Bukanov, Nickolai O.; Westblom, T. Ulf
  • Nucleic Acids Research, Vol. 20, Issue 19
  • DOI: 10.1093/nar/20.19.5137

Effect of pH and lactic or acetic acid on ethanol productivity by Saccharomyces cerevisiae in corn mash
journal, February 2006

  • Graves, Tara; Narendranath, Neelakantam V.; Dawson, Karl
  • Journal of Industrial Microbiology & Biotechnology, Vol. 33, Issue 6
  • DOI: 10.1007/s10295-006-0091-6

Bacterial contaminants of fuel ethanol production
journal, August 2004

  • Skinner, Kelly A.; Leathers, Timothy D.
  • Journal of Industrial Microbiology & Biotechnology, Vol. 31, Issue 9
  • DOI: 10.1007/s10295-004-0159-0

Death by a thousand cuts: the challenges and diverse landscape of lignocellulosic hydrolysate inhibitors
journal, March 2014


Screening of Saccharomyces cerevisiae strains with respect to anaerobic growth in non-detoxified lignocellulose hydrolysate
journal, July 2009


Short-term adaptation improves the fermentation performance of Saccharomyces cerevisiae in the presence of acetic acid at low pH
journal, July 2013

  • Sànchez i. Nogué, Violeta; Narayanan, Venkatachalam; Gorwa-Grauslund, Marie F.
  • Applied Microbiology and Biotechnology, Vol. 97, Issue 16
  • DOI: 10.1007/s00253-013-5093-5

Fermentation of lignocellulosic hydrolysates. II: inhibitors and mechanisms of inhibition
journal, August 2000


Metabolomic Study of Interactive Effects of Phenol, Furfural, and Acetic Acid on Saccharomyces cerevisiae
journal, October 2011

  • Ding, Ming-Zhu; Wang, Xin; Yang, Yang
  • OMICS: A Journal of Integrative Biology, Vol. 15, Issue 10
  • DOI: 10.1089/omi.2011.0003

Stochastic switching as a survival strategy in fluctuating environments
journal, March 2008

  • Acar, Murat; Mettetal, Jerome T.; van Oudenaarden, Alexander
  • Nature Genetics, Vol. 40, Issue 4
  • DOI: 10.1038/ng.110

Importance of glucose-6-phosphate dehydrogenase (G6PDH) for vanillin tolerance in Saccharomyces cerevisiae
journal, September 2014

  • Nguyen, Trinh Thi My; Kitajima, Sakihito; Izawa, Shingo
  • Journal of Bioscience and Bioengineering, Vol. 118, Issue 3
  • DOI: 10.1016/j.jbiosc.2014.02.025

Polygenic analysis and targeted improvement of the complex trait of high acetic acid tolerance in the yeast Saccharomyces cerevisiae
journal, January 2016

  • Meijnen, Jean-Paul; Randazzo, Paola; Foulquié-Moreno, María R.
  • Biotechnology for Biofuels, Vol. 9, Issue 1
  • DOI: 10.1186/s13068-015-0421-x

Enhanced resistance of Saccharomyces cerevisiae to vanillin by expression of lacA from Trametes sp. AH28-2
journal, September 2011


A 5-hydroxymethyl furfural reducing enzyme encoded by theSaccharomyces cerevisiae ADH6 gene conveys HMF tolerance
journal, January 2006

  • Petersson, Anneli; Almeida, João R. M.; Modig, Tobias
  • Yeast, Vol. 23, Issue 6, p. 455-464
  • DOI: 10.1002/yea.1370

Isolation and characterization of a resident tolerant Saccharomyces cerevisiae strain from a spent sulfite liquor fermentation plant
journal, January 2012

  • Sànchez i. Nogué, Violeta; Bettiga, Maurizio; Gorwa-Grauslund, Marie F.
  • AMB Express, Vol. 2, Issue 1
  • DOI: 10.1186/2191-0855-2-68

Lignocellulosic biomass for bioethanol production: Current perspectives, potential issues and future prospects
journal, August 2012


Modifying Yeast Tolerance to Inhibitory Conditions of Ethanol Production Processes
journal, November 2015

  • Caspeta, Luis; Castillo, Tania; Nielsen, Jens
  • Frontiers in Bioengineering and Biotechnology, Vol. 3
  • DOI: 10.3389/fbioe.2015.00184

Modeling bacterial contamination of fuel ethanol fermentation
journal, May 2009

  • Bischoff, Kenneth M.; Liu, Siqing; Leathers, Timothy D.
  • Biotechnology and Bioengineering, Vol. 103, Issue 1
  • DOI: 10.1002/bit.22244

Yeast population dynamics of industrial fuel-ethanol fermentation process assessed by PCR-fingerprinting
journal, August 2005

  • Silva-Filho, Eurípedes Alves da; Santos, Scheila Karina Brito dos; Resende, Alecsandra do Monte
  • Antonie van Leeuwenhoek, Vol. 88, Issue 2
  • DOI: 10.1007/s10482-005-7283-3

    Works referencing / citing this record:

    Replacing water and nutrients for ethanol production by ARTP derived biogas slurry tolerant Zymomonas mobilis strain
    journal, May 2019


    Replacing water and nutrients for ethanol production by ARTP derived biogas slurry tolerant Zymomonas mobilis strain
    journal, May 2019


    Overcoming lignocellulose‐derived microbial inhibitors: advancing the Saccharomyces cerevisiae resistance toolbox
    journal, August 2019

    • Brandt, Bianca A.; Jansen, Trudy; Görgens, Johann F.
    • Biofuels, Bioproducts and Biorefining, Vol. 13, Issue 6
    • DOI: 10.1002/bbb.2042

    Repeated cultures of Saccharomyces cerevisiae SC90 to tolerate inhibitors generated during cassava processing waste hydrolysis for bioethanol production
    journal, February 2019

    • Palakawong Na Ayutthaya, Pakathamon; Charoenrat, Theppanya; Krusong, Warawut
    • 3 Biotech, Vol. 9, Issue 3
    • DOI: 10.1007/s13205-019-1607-x

    Increased lignocellulosic inhibitor tolerance of Saccharomyces cerevisiae cell populations in early stationary phase
    journal, May 2017

    • Narayanan, Venkatachalam; Schelin, Jenny; Gorwa-Grauslund, Marie
    • Biotechnology for Biofuels, Vol. 10, Issue 1
    • DOI: 10.1186/s13068-017-0794-0

    Combined ensiling and hydrothermal processing as efficient pretreatment of sugarcane bagasse for 2G bioethanol production
    journal, December 2018

    • Ambye-Jensen, Morten; Balzarotti, Riccardo; Thomsen, Sune Tjalfe
    • Biotechnology for Biofuels, Vol. 11, Issue 1
    • DOI: 10.1186/s13068-018-1338-y

    Engineered Zymomonas mobilis tolerant to acetic acid and low pH via multiplex atmospheric and room temperature plasma mutagenesis
    journal, January 2019


      Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.