Abstract
Furfural and 5-hydroxymethylfurfural (HMF) are representative inhibitors generated from biomass pretreatment using dilute acid hydrolysis that interfere with yeast growth and subsequent fermentation. Few yeast strains tolerant to inhibitors are available. In this study, we report a tolerant strain, Saccharomyces cerevisiae NRRL Y-50049, which has enhanced biotransformation ability to convert furfural to furan methanol (FM), HMF to furan di-methanol (FDM), and produce a normal yield of ethanol. Our recent identification of HMF and development of protocol to synthesize the HMF metabolic conversion product FDM allowed studies on fermentation metabolic kinetics in the presence of HMF and furfural. Individual gene-encoding enzymes possessing aldehyde reduction activities demonstrated cofactor preference for NADH or NADPH. However, protein extract from whole yeast cells showed equally strong aldehyde reduction activities coupled with either cofactor. Deletion of a single candidate gene did not affect yeast growth in the presence of the inhibitors. Our results suggest that detoxification of furfural and HMF by the ethanologenic yeast S. cerevisiae strain Y-50049 likely involves multiple gene mediated NAD(P)H-dependent aldehyde reduction. Conversion pathways of furfural and HMF relevant to glycolysis and ethanol production were refined based on our findings in this study.
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Acknowledgments
We are grateful for technical assistance contributed by Amy Cash, Suzanne Milborne, and Maureen Shea-Andersh. This study was supported by US Department of Agriculture ARS National Programs 307/306 and, in part, supported by USDA HQ Mentor of Postdoctoral Research Associate Program and the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2006-35504-17359 to ZLL.
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Lewis Liu, Z., Moon, J., Andersh, B.J. et al. Multiple gene-mediated NAD(P)H-dependent aldehyde reduction is a mechanism of in situ detoxification of furfural and 5-hydroxymethylfurfural by Saccharomyces cerevisiae . Appl Microbiol Biotechnol 81, 743–753 (2008). https://doi.org/10.1007/s00253-008-1702-0
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DOI: https://doi.org/10.1007/s00253-008-1702-0