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Engineering and two-stage evolution of a lignocellulosic hydrolysate-tolerant Saccharomyces cerevisiae strain for anaerobic fermentation of xylose from AFEX pretreated corn stover

Parreiras, Lucas S. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Breuer, Rebecca J. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Narasimhan, Ragothaman Avanasi (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
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Higbee, Alan J. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
La Reau, Alex (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Tremaine, Mary T. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Qin, Li (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Willis, Laura B. (author)
Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Bice, Benjamin D. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Bonfert, Brandi L. (author)
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Pinhancos, Rebeca C. (author)
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Balloon, Allison J. (author)
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Uppugundla, Nirmal (author)
DOE Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, Michigan, United States of America; Biomass Conversion Research Laboratory, Department of Chemical Engineering and Materials Science, Michigan State University, Lansing, Michigan, United States of America
Liu, Tongjun (author)
DOE Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, Michigan, United States of America; School of Food and Bioengineering, Qilu University of Technology, Jinan, China
Li, Chenlin (author)
Advanced Biofuels Process Demonstration Unit, Lawrence Berkeley National Laboratory, Emeryville, California, United States of America
Tanjore, Deepti (author)
Advanced Biofuels Process Demonstration Unit, Lawrence Berkeley National Laboratory, Emeryville, California, United States of America
Ong, Irene (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Li, Haibo (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Pohlmann, Edward L. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Serate, Jose (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Withers, Sydnor T. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Simmons, Blake Alexander (author)
Deconstruction Division, Joint BioEnergy Institute, Emeryville, California, United States of America
Hodge, David (author)
Luleå tekniska universitet,Industriell miljö- och processteknik,DOE Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, Michigan, United States of America; Department of Chemical Engineering & Materials Science, Michigan State University, East Lansing, Michigan, United States of America; Department of Biosystems & Agricultural Engineering, Michigan State University, East Lansing Michigan, United States of America
Westphall, Michael S. (author)
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Coon, Joshua J. (author)
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Dale, Bruce E. (author)
DOE Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, Michigan, United States of America; Biomass Conversion Research Laboratory, Department of Chemical Engineering and Materials Science, Michigan State University, Lansing, Michigan, United States of America
Balan, Venkatesh (author)
DOE Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, Michigan, United States of America; Biomass Conversion Research Laboratory, Department of Chemical Engineering and Materials Science, Michigan State University, Lansing, Michigan, United States of America
Keating, David H. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Zhang, Yaoping (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Landick, Robert (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Gasch, Audrey P. (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Laboratory of Genetics, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
Sato, Trey (author)
DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America
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 (creator_code:org_t)
2014-09-15
2014
English.
In: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 9:9
Related links:
https://doi.org/10.1...
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  • Journal article (peer-reviewed)
Abstract Subject headings
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  • The inability of the yeast Saccharomyces cerevisiae to ferment xylose effectively under anaerobic conditions is a major barrier to economical production of lignocellulosic biofuels. Although genetic approaches have enabled engineering of S. cerevisiae to convert xylose efficiently into ethanol in defined lab medium, few strains are able to ferment xylose from lignocellulosic hydrolysates in the absence of oxygen. This limited xylose conversion is believed to result from small molecules generated during biomass pretreatment and hydrolysis, which induce cellular stress and impair metabolism. Here, we describe the development of a xylose-fermenting S. cerevisiae strain with tolerance to a range of pretreated and hydrolyzed lignocellulose, including Ammonia Fiber Expansion (AFEX)-pretreated corn stover hydrolysate (ACSH). We genetically engineered a hydrolysate-resistant yeast strain with bacterial xylose isomerase and then applied two separate stages of aerobic and anaerobic directed evolution. The emergent S. cerevisiae strain rapidly converted xylose from lab medium and ACSH to ethanol under strict anaerobic conditions. Metabolomic, genetic and biochemical analyses suggested that a missense mutation in GRE3, which was acquired during the anaerobic evolution, contributed toward improved xylose conversion by reducing intracellular production of xylitol, an inhibitor of xylose isomerase. These results validate our combinatorial approach, which utilized phenotypic strain selection, rational engineering and directed evolution for the generation of a robust S. cerevisiae strain with the ability to ferment xylose anaerobically from ACSH.

Subject headings

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Bioprocessteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Bioprocess Technology (hsv//eng)

Keyword

Biokemisk processteknik
Biochemical Process Engineering

Publication and Content Type

ref (subject category)
art (subject category)

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