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Title: Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading

Abstract

Lignocellulosic biomass is an abundant, renewable feedstock useful for the production of fuel-grade ethanol via the processing steps of pretreatment, enzyme hydrolysis, and microbial fermentation. Traditional industrial yeasts do not ferment xylose and are not able to grow, survive, or ferment in concentrated hydrolyzates that contain enough sugar to support economical ethanol recovery since they are laden with toxic byproducts generated during pretreatment. Repetitive culturing in two types of concentrated hydrolyzates was applied along with ethanol challenged xylose-fed continuous culture to force targeted evolution of the native pentose fermenting yeast Scheffersomyces (Pichia) stipitis strain NRRL Y-7124 maintained in the ARS Culture Collection, Peoria, IL. Isolates collected from various enriched populations were screened and ranked based on relative xylose uptake rate and ethanol yield. Ranking on hydrolyzates with and without nutritional supplementation was used to identify those isolates with best performance across diverse conditions. Robust S. stipitis strains adapted to perform very well in enzyme hydrolyzates of high solids loading ammonia fiber expansion-pretreated corn stover (18% weight per volume solids) and dilute sulfuric acid-pretreated switchgrass (20% w/v solids) were obtained. Improved features include reduced initial lag phase preceding growth, significantly enhanced fermentation rates, improved ethanol tolerance and yield, reduced diauxic lagmore » during glucose-xylose transition, and ability to accumulate >40 g/L ethanol in <167 h when fermenting hydrolyzate at low initial cell density of 0.5 absorbance units and pH 5 to 6.« less

Authors:
 [1];  [1];  [1];  [1];  [2];  [3];  [3];  [3];  [3];  [1]
+ Show Author Affiliations
  1. USDA National Center for Agricultural Utilization Research, Peoria, IL (United States). Bioenergy Research Unit.
  2. USDA National Center for Agricultural Utilization Research, Peoria, IL (United States). Bacterial Foodborne Pathogens and Mycology Research.
  3. Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Research Center (GLBRC).
Publication Date:
Research Org.:
Great Lakes Bioenergy Research Center (GLBRC), Michigan State Univ., East Lansing, MI (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1184797
Grant/Contract Number:  
FC02-07ER64494
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; lignocellulose; biofuel; adaptation; yeast; Pichia stipitis; fermentation

Citation Formats

Slininger, Patricia J., Shea-Andersh, Maureen A., Thompson, Stephanie R., Dien, Bruce S., Kurtzman, Cletus P., Balan, Venkatesh, da Costa Sousa, Leonardo, Uppugundla, Nirmal, Dale, Bruce E, and Cotta, Michael A. Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading. United States: N. p., 2015. Web. doi:10.1186/s13068-015-0239-6.
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Slininger, Patricia J., Shea-Andersh, Maureen A., Thompson, Stephanie R., Dien, Bruce S., Kurtzman, Cletus P., Balan, Venkatesh, da Costa Sousa, Leonardo, Uppugundla, Nirmal, Dale, Bruce E, & Cotta, Michael A. Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading. United States. https://doi.org/10.1186/s13068-015-0239-6
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Slininger, Patricia J., Shea-Andersh, Maureen A., Thompson, Stephanie R., Dien, Bruce S., Kurtzman, Cletus P., Balan, Venkatesh, da Costa Sousa, Leonardo, Uppugundla, Nirmal, Dale, Bruce E, and Cotta, Michael A. Thu . "Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading". United States. https://doi.org/10.1186/s13068-015-0239-6. https://www.osti.gov/servlets/purl/1184797.
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@article{osti_1184797,
title = {Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading},
author = {Slininger, Patricia J. and Shea-Andersh, Maureen A. and Thompson, Stephanie R. and Dien, Bruce S. and Kurtzman, Cletus P. and Balan, Venkatesh and da Costa Sousa, Leonardo and Uppugundla, Nirmal and Dale, Bruce E and Cotta, Michael A},
abstractNote = {Lignocellulosic biomass is an abundant, renewable feedstock useful for the production of fuel-grade ethanol via the processing steps of pretreatment, enzyme hydrolysis, and microbial fermentation. Traditional industrial yeasts do not ferment xylose and are not able to grow, survive, or ferment in concentrated hydrolyzates that contain enough sugar to support economical ethanol recovery since they are laden with toxic byproducts generated during pretreatment. Repetitive culturing in two types of concentrated hydrolyzates was applied along with ethanol challenged xylose-fed continuous culture to force targeted evolution of the native pentose fermenting yeast Scheffersomyces (Pichia) stipitis strain NRRL Y-7124 maintained in the ARS Culture Collection, Peoria, IL. Isolates collected from various enriched populations were screened and ranked based on relative xylose uptake rate and ethanol yield. Ranking on hydrolyzates with and without nutritional supplementation was used to identify those isolates with best performance across diverse conditions. Robust S. stipitis strains adapted to perform very well in enzyme hydrolyzates of high solids loading ammonia fiber expansion-pretreated corn stover (18% weight per volume solids) and dilute sulfuric acid-pretreated switchgrass (20% w/v solids) were obtained. Improved features include reduced initial lag phase preceding growth, significantly enhanced fermentation rates, improved ethanol tolerance and yield, reduced diauxic lag during glucose-xylose transition, and ability to accumulate >40 g/L ethanol in <167 h when fermenting hydrolyzate at low initial cell density of 0.5 absorbance units and pH 5 to 6.},
doi = {10.1186/s13068-015-0239-6},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 8,
place = {United States},
year = {Thu Apr 09 00:00:00 EDT 2015},
month = {Thu Apr 09 00:00:00 EDT 2015}
}
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https://doi.org/10.1186/s13068-015-0239-6
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  • Applied Microbiology and Biotechnology, Vol. 72, Issue 6
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Adaptive response of yeasts to furfural and 5-hydroxymethylfurfural and new chemical evidence for HMF conversion to 2,5-bis-hydroxymethylfuran
journal, July 2004

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Stoichiometry and Kinetics of Xylose Fermentation by Pichia stipitis
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Dehydration of Ethanol: New Approach Gives Positive Energy Balance
journal, August 1979

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    Works referencing / citing this record:

    Adaptive laboratory evolution of nanocellulose‐producing bacterium
    journal, May 2019

    • Vasconcellos, Vanessa M.; Farinas, Cristiane S.; Ximenes, Eduardo
    • Biotechnology and Bioengineering, Vol. 116, Issue 8
    • DOI: 10.1002/bit.26997

    Synthetic biology for evolutionary engineering: from perturbation of genotype to acquisition of desired phenotype
    journal, May 2019

    Genomic and transcriptomic analysis of Candida intermedia reveals the genetic determinants for its xylose-converting capacity
    journal, March 2020

    • Geijer, Cecilia; Faria-Oliveira, Fábio; Moreno, Antonio D.
    • Biotechnology for Biofuels, Vol. 13, Issue 1
    • DOI: 10.1186/s13068-020-1663-9

    Global Warming Potential of Biomass-to-Ethanol: Review and Sensitivity Analysis through a Case Study
    journal, July 2019

    Genome and metabolic engineering in non-conventional yeasts: Current advances and applications
    journal, September 2017

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