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Applied Microbiology and Biotechnology

, Volume 97, Issue 11, pp 4811–4819 | Cite as

Utilization of Saccharomyces cerevisiae recombinant strain incapable of both ethanol and glycerol biosynthesis for anaerobic bioproduction

  • Yoshihiro Ida
  • Takashi Hirasawa
  • Chikara Furusawa
  • Hiroshi ShimizuEmail author
Biotechnological products and process engineering

Abstract

The yeast Saccharomyces cerevisiae produces ethanol and glycerol as major unwanted byproducts, unless ethanol and glycerol are the target compounds. Minimizing the levels of these byproducts is important for bioproduction processes using yeast cells. In this study, we constructed a yeast strain in which both ethanol and glycerol production pathways were disrupted and examined its culture characteristics. In wild-type yeast strain, metabolic pathways that produce ethanol and glycerol play an important role in reoxidizing nicotinamide adenine dinucleotide (NADH) generated during glycolysis, particularly under anaerobic conditions. Strains in which both pathways were disrupted therefore failed to grow and consume glucose under anaerobic conditions. Introduction of desired metabolic reaction(s) coupled with NADH oxidation enabled the engineered strain to consume substrate and produce target compound(s). Here we introduced NADH-oxidization-coupled L-lactate production mechanisms into a yeast strain incapable of ethanol and glycerol biosynthesis, based on in silico simulation using a genome-scale metabolic model of S. cerevisiae. From the results of in silico simulation based on flux balance analysis, a feasible anaerobic non-growing metabolic state, in which L-lactate yield approached the theoretical maximum, was identified and this phenomenon was verified experimentally. The yeast strain incapable of both ethanol and glycerol biosynthesis is a potentially valuable host for bioproduction coupled with NADH oxidation under anaerobic conditions.

Keywords

Saccharomyces cerevisiae Bioproduction Ethanol Glycerol Lactate Genome-scale metabolic model 

Notes

Acknowledgments

The plasmid pNV11 was provided by the National Bio-Resource Project (Yeast) of the Ministry of Education, Culture, Sports, Science, and Technology of Japan. This work was financially supported by a Grant-in-Aid for Scientific Research (A) to SH (no. 24246134) from the Japan Society for Promotion of Science. This work was also supported in part by the Global COE Program of the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

Supplementary material

253_2013_4760_MOESM1_ESM.pdf (902 kb)
ESM 1 (PDF 901 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Yoshihiro Ida
    • 1
  • Takashi Hirasawa
    • 1
  • Chikara Furusawa
    • 1
    • 2
  • Hiroshi Shimizu
    • 1
    Email author
  1. 1.Department of Bioinformatic Engineering, Graduate School of Information Science and TechnologyOsaka UniversitySuitaJapan
  2. 2.Quantitative Biology CenterRIKENSuitaJapan

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