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Enhanced xylose fermentation by engineered yeast expressing NADH oxidase through high cell density inoculums

  • Fermentation, Cell Culture and Bioengineering - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Accumulation of reduced byproducts such as glycerol and xylitol during xylose fermentation by engineered Saccharomyces cerevisiae hampers the economic production of biofuels and chemicals from cellulosic hydrolysates. In particular, engineered S. cerevisiae expressing NADPH-linked xylose reductase (XR) and NAD+-linked xylitol dehydrogenase (XDH) produces substantial amounts of the reduced byproducts under anaerobic conditions due to the cofactor difference of XR and XDH. While the additional expression of a water-forming NADH oxidase (NoxE) from Lactococcus lactis in engineered S. cerevisiae with the XR/XDH pathway led to reduced glycerol and xylitol production and increased ethanol yields from xylose, volumetric ethanol productivities by the engineered yeast decreased because of growth defects from the overexpression of noxE. In this study, we introduced noxE into an engineered yeast strain (SR8) exhibiting near-optimal xylose fermentation capacity. To overcome the growth defect caused by the overexpression of noxE, we used a high cell density inoculum for xylose fermentation by the SR8 expressing noxE. The resulting strain, SR8N, not only showed a higher ethanol yield and lower byproduct yields, but also exhibited a high ethanol productivity during xylose fermentation. As noxE overexpression elicits a negligible growth defect on glucose conditions, the beneficial effects of noxE overexpression were substantial when a mixture of glucose and xylose was used. Consumption of glucose led to rapid cell growth and therefore enhanced the subsequent xylose fermentation. As a result, the SR8N strain produced more ethanol and fewer byproducts from a mixture of glucose and xylose than the parental SR8 strain without noxE overexpression. Our results suggest that the growth defects from noxE overexpression can be overcome in the case of fermenting lignocellulose-derived sugars such as glucose and xylose.

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Acknowledgements

This work was supported by funding from the Energy Biosciences Institute and TLT would like to acknowledge that this project was supported by the Agriculture and Food Research Initiative Competitive Grant No. 2015-67011-22806 from the USDA National Institute of Food and Agriculture.

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  1. Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL, 61801, USA

    Guo-Chang Zhang & Yong-Su Jin

  2. Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Guo-Chang Zhang, Timothy L. Turner & Yong-Su Jin

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  1. Guo-Chang Zhang
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  2. Timothy L. Turner
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Correspondence to Yong-Su Jin.

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Zhang, GC., Turner, T.L. & Jin, YS. Enhanced xylose fermentation by engineered yeast expressing NADH oxidase through high cell density inoculums. J Ind Microbiol Biotechnol 44, 387–395 (2017). https://doi.org/10.1007/s10295-016-1899-3

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