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Applied Microbiology and Biotechnology

, Volume 81, Issue 2, pp 243–255 | Cite as

Expression of protein engineered NADP+-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae

  • Akinori MatsushikaEmail author
  • Seiya Watanabe
  • Tsutomu Kodaki
  • Keisuke Makino
  • Hiroyuki Inoue
  • Katsuji Murakami
  • Osamu Takimura
  • Shigeki Sawayama
Biotechnological Products and Process Engineering

Abstract

A recombinant Saccharomyces cerevisiae strain transformed with xylose reductase (XR) and xylitol dehydrogenase (XDH) genes from Pichia stipitis has the ability to convert xylose to ethanol together with the unfavorable excretion of xylitol, which may be due to cofactor imbalance between NADPH-preferring XR and NAD+-dependent XDH. To reduce xylitol formation, we have already generated several XDH mutants with a reversal of coenzyme specificity toward NADP+. In this study, we constructed a set of recombinant S. cerevisiae strains with xylose-fermenting ability, including protein-engineered NADP+-dependent XDH-expressing strains. The most positive effect on xylose-to-ethanol fermentation was found by using a strain named MA-N5, constructed by chromosomal integration of the gene for NADP+-dependent XDH along with XR and endogenous xylulokinase genes. The MA-N5 strain had an increase in ethanol production and decrease in xylitol excretion compared with the reference strain expressing wild-type XDH when fermenting not only xylose but also mixed sugars containing glucose and xylose. Furthermore, the MA-N5 strain produced ethanol with a high yield of 0.49 g of ethanol/g of total consumed sugars in the nonsulfuric acid hydrolysate of wood chips. The results demonstrate that glucose and xylose present in the lignocellulosic hydrolysate can be efficiently fermented by this redox-engineered strain.

Keywords

Xylitol dehydrogenase Coenzyme specificity Saccharomyces cerevisiae Xylose fermentation Ethanol production 

Notes

Acknowledgments

We thank Ms. Maiko Kato for her technical support. This study was supported by the New Energy and Industrial Technology Development Organization (NEDO), Japan.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Akinori Matsushika
    • 1
    Email author
  • Seiya Watanabe
    • 2
  • Tsutomu Kodaki
    • 2
  • Keisuke Makino
    • 2
  • Hiroyuki Inoue
    • 1
  • Katsuji Murakami
    • 1
  • Osamu Takimura
    • 1
  • Shigeki Sawayama
    • 1
  1. 1.Biomass Technology Research CenterNational Institute of Advanced Industrial Science and TechnologyKureJapan
  2. 2.Institute of Advanced EnergyKyoto UniversityUjiJapan

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