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A genetic overhaul of Saccharomyces cerevisiae 424A(LNH-ST) to improve xylose fermentation

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Robust microorganisms are necessary for economical bioethanol production. However, such organisms must be able to effectively ferment both hexose and pentose sugars present in lignocellulosic hydrolysate to ethanol. Wild type Saccharomyces cerevisiae can rapidly ferment hexose, but cannot ferment pentose sugars. Considerable efforts were made to genetically engineer S. cerevisiae to ferment xylose. Our genetically engineered S cerevisiae yeast, 424A(LNH-ST), expresses NADPH/NADH xylose reductase (XR) that prefer NADPH and NAD+-dependent xylitol dehydrogenase (XD) from Pichia stipitis, and overexpresses endogenous xylulokinase (XK). This strain is able to ferment glucose and xylose, as well as other hexose sugars, to ethanol. However, the preference for different cofactors by XR and XD might lead to redox imbalance, xylitol excretion, and thus might reduce ethanol yield and productivity. In the present study, genes responsible for the conversion of xylose to xylulose with different cofactor specificity (1) XR from N. crassa (NADPH-dependent) and C. parapsilosis (NADH-dependent), and (2) mutant XD from P. stipitis (containing three mutations D207A/I208R/F209S) were overexpressed in wild type yeast. To increase the NADPH pool, the fungal GAPDH enzyme from Kluyveromyces lactis was overexpressed in the 424A(LNH-ST) strain. Four pentose phosphate pathway (PPP) genes, TKL1, TAL1, RKI1 and RPE1 from S. cerevisiae, were also overexpressed in 424A(LNH-ST). Overexpression of GAPDH lowered xylitol production by more than 40%. However, other strains carrying different combinations of XR and XD, as well as new strains containing the overexpressed PPP genes, did not yield any significant improvement in xylose fermentation.

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Acknowledgments

The project was financially supported by the US Department of Energy Biomass Program, Contract G017059-16649.

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Authors and Affiliations

  1. Laboratory of Renewable Resources Engineering, Purdue University, West Lafayette, IN, 47907, USA

    Aloke K. Bera, Nancy W. Y. Ho, Aftab Khan & Miroslav Sedlak

  2. School of Chemical Engineering, Purdue University, West Lafayette, IN, 47907, USA

    Nancy W. Y. Ho

  3. Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN, 47907, USA

    Miroslav Sedlak

Authors
  1. Aloke K. Bera
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  2. Nancy W. Y. Ho
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  3. Aftab Khan
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  4. Miroslav Sedlak
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Correspondence to Miroslav Sedlak.

Additional information

This article is based on a presentation at the 32nd Symposium on Biotechnology for Fuels and Chemicals.

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Bera, A.K., Ho, N.W.Y., Khan, A. et al. A genetic overhaul of Saccharomyces cerevisiae 424A(LNH-ST) to improve xylose fermentation. J Ind Microbiol Biotechnol 38, 617–626 (2011). https://doi.org/10.1007/s10295-010-0806-6

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  • DOI: https://doi.org/10.1007/s10295-010-0806-6

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