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ARS Home » Midwest Area » Peoria, Illinois » National Center for Agricultural Utilization Research » Renewable Product Technology Research » Research » Publications at this Location » Publication #262617
Title: Random UV-C mutagenesis of Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 to improve anaerobic growth on lignocellulosic sugars

Author
item Hughes, Stephen
item Bischoff, Kenneth
item GIBBONS, WILLIAM - South Dakota State University
item BANG, SOOKIE - South Dakota School Of Mines And Technology
item PINKELMAN, REBECCA - South Dakota School Of Mines And Technology
item Slininger, Patricia - Pat
item Qureshi, Nasib
item Liu, Siqing
item Saha, Badal
item JACKSON, JOHN - Former ARS Employee
item Cotta, Michael
item Rich, Joseph
item JAVERS, JEREMY - Icm, Inc

Submitted to: Journal of Industrial Microbiology and Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/15/2011
Publication Date: 1/1/2012
Citation: Hughes, S.R., Bischoff, K.M., Gibbons, W.R., Bang, S.S., Pinkelman, R., Slininger, P.J., Qureshi, N., Liu, S., Saha, B.C., Jackson, J.S., Cotta, M.A., Rich, J.O., Javers, J. 2012. Random UV-C mutagenesis of Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 to improve anaerobic growth on lignocellulosic sugars. Journal of Industrial Microbiology and Biotechnology. 39(1):163–173.

Interpretive Summary: Yeast strains for anaerobic conversion of five- and six-carbon sugars to ethanol were produced from Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 by using UV-C irradiation that produced mutations in the genome and generate unique strains. The irradiated strains were capable of growing anaerobically on xylose plus glucose as the carbon sources with higher ethanol production than a Saccharomyces cerevisiae yeast strain during 250 to 500-hour growth. These new strains have potential application in industrial high-yield fuel ethanol production from plant biomass.

Technical Abstract: Yeast strains for anaerobic conversion of lignocellulosic sugars to ethanol were produced from Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 using UV-C mutagenesis. Random UV-C mutagenesis potentially produces large numbers of mutations broadly and uniformly over the whole genome to generate unique strains. Wild-type cultures of S. stipitis NRRL Y-7124 were subjected to 4-hour irradiation with UV-C (234 nm) targeted to kill approximately 60% of cells. When surviving cells were selected in sufficient numbers via automated plating strategies and cultured anaerobically on xylose medium for 5 months at 28 deg C, five novel mutagenized S. stipitis strains were obtained. It was determined using variable number tandem repeat (VNTR) analysis that mutations had occurred in the genome, which may have produced genes that allowed the anaerobic utilization of xylose. The mutagenized strains were capable of growing anaerobically on xylose/glucose substrate with higher ethanol production than a Saccharomyces cerevisiae yeast strain during 250 to 500-hour growth. The Scheffersomyces stipitis strains resulting from this intense multigene mutagenesis strategy have potential application in industrial high-yield fuel ethanol production anaerobically for fermentation of lignocellulosic hydrolysates.