Tolerance to furfural-induced stress is associated with pentose phosphate pathway genes ZWF1, GND1, RPE1, and TKL1 in Saccharomyces cerevisiae

Engineering yeast to be more tolerant to fermentation inhibitors, furfural and 5-hydroxymethylfurfural (HMF), will lead to more efficient lignocellulose to ethanol bioconversion. To identify target genes involved in furfural tolerance, a Saccharomyces cerevisiae gene disruption library was screened...

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Published in:Applied microbiology and biotechnology 2006-07, Vol.71 (3), p.339-349
Main Authors: GORSICH, S. W, DIEN, B. S, NICHOLS, N. N, SLININGER, P. J, LIU, Z. L, SKORY, C. D
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Carbohydrate Epimerases - genetics
Carbohydrate Epimerases - metabolism
Cellulose - metabolism
Fundamental and applied biological sciences. Psychology
Furaldehyde - analogs & derivatives
Furaldehyde - pharmacology
Gene Expression Regulation, Fungal
Genetics
Glucosephosphate Dehydrogenase - genetics
Glucosephosphate Dehydrogenase - metabolism
Heat-Shock Response
Lignin - metabolism
Mutation
Pentose Phosphate Pathway
Proteins
Saccharomyces cerevisiae
Saccharomyces cerevisiae - drug effects
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - growth & development
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
Transketolase - genetics
Transketolase - metabolism
Yeast
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Summary:Engineering yeast to be more tolerant to fermentation inhibitors, furfural and 5-hydroxymethylfurfural (HMF), will lead to more efficient lignocellulose to ethanol bioconversion. To identify target genes involved in furfural tolerance, a Saccharomyces cerevisiae gene disruption library was screened for mutants with growth deficiencies in the presence of furfural. It was hypothesized that overexpression of these genes would provide a growth benefit in the presence of furfural. Sixty two mutants were identified whose corresponding genes function in a wide spectrum of physiological pathways, suggesting that furfural tolerance is a complex process. We focused on four mutants, zwf1, gnd1, rpe1, and tkl1, which represent genes encoding pentose phosphate pathway (PPP) enzymes. At various concentrations of furfural and HMF, a clear association with higher sensitivity to these inhibitors was demonstrated in these mutants. PPP mutants were inefficient at reducing furfural to the less toxic furfuryl alcohol, which we propose is a result of an overall decreased abundance of reducing equivalents or to NADPH's role in stress tolerance. Overexpression of ZWF1 in S. cerevisiae allowed growth at furfural concentrations that are normally toxic. These results demonstrate a strong relationship between PPP genes and furfural tolerance and provide additional putative target genes involved in furfural tolerance.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-005-0142-3