NDLI: Engineering and Two-Stage Evolution of a Lignocellulosic Hydrolysate-Tolerant Saccharomyces cerevisiae Strain for Anaerobic Fermentation of Xylose from AFEX Pretreated Corn Stover

Content Provider	PubMed Central
Author	Parreiras, Lucas S. Breuer, Rebecca J. Narasimhan, Ragothaman Avanasi Higbee, Alan J. Reau, Alex La Tremaine, Mary Qin, Li Willis, Laura B. Bice, Benjamin D. Bonfert, Brandi L. Pinhancos, Rebeca C. Balloon, Allison J. Nirmal, Uppugundla Liu, Tongjun Li, Chenlin Deepti, Tanjore Ong, Irene M. Haibo, Li Pohlmann, Edward L. Jose, Serate Withers, Sydnor T. Simmons, Blake A. Hodge, David B. Westphall, Michael S. Coon, Joshua J. Dale, Bruce E. Balan, Venkatesh Keating, David H. Zhang, Yaoping Landick, Robert Gasch, Audrey P. Sato, Trey K.
Editor	Zhang, Y-h Percival
Copyright Year	2014
Abstract	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.
Related Links	http://dx.doi.org/10.1371/journal.pone.0107499
Starting Page	107499
File Format	PDF
ISSN	19326203
e-ISSN	19326203
Journal	PLoS ONE
Issue Number	9
Volume Number	9
Language	English
Publisher	Public Library of Science
Publisher Date	2014-09-01
Access Restriction	Open
Rights Holder	Public Library of Science
Subject Keyword	Biochemistry, Genetics and Molecular Biology(all) Agricultural and Biological Sciences(all) Medicine(all) Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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