¹³C-based metabolic flux analysis of Saccharomyces cerevisiae with a reduced Crabtree effect

Shuichi Kajihata; Fumio Matsuda; Mika Yoshimi; Kenshi Hayakawa; Chikara Furusawa; Akihisa Kanda; Hiroshi Shimizu

doi:10.1016/j.jbiosc.2014.12.014

¹³C-based metabolic flux analysis of Saccharomyces cerevisiae with a reduced Crabtree effect

J Biosci Bioeng. 2015 Aug;120(2):140-4. doi: 10.1016/j.jbiosc.2014.12.014. Epub 2015 Jan 26.

Authors

Shuichi Kajihata¹, Fumio Matsuda², Mika Yoshimi³, Kenshi Hayakawa⁴, Chikara Furusawa⁵, Akihisa Kanda⁶, Hiroshi Shimizu⁷

Affiliations

¹ Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: s-kajihata@ist.osaka-u.ac.jp.
² Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: fmatsuda@ist.osaka-u.ac.jp.
³ Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: mika_yoshimi@bio.eng.osaka-u.ac.jp.
⁴ KANEKA Fundamental Technology Research Alliance Laboratories, Graduate School of Engineering, Osaka University, 2-8 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: Kenshi.Hayakawa@kaneka.co.jp.
⁵ Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan; Quantitative Biology Center, RIKEN, 6-2-3 Furuedai, Suita, Osaka 565-0874, Japan. Electronic address: chikara.furusawa@riken.jp.
⁶ New & Fundamental Technology Group Process Technology Laboratories, KANEKA Corporation, 1-8 Miyamae-cho, Takasago-cho, Takasago, Hyogo 676-8688, Japan. Electronic address: Akihisa_Kanda@kn.kaneka.co.jp.
⁷ Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: shimizu@ist.osaka-u.ac.jp.

PMID: 25634548
DOI: 10.1016/j.jbiosc.2014.12.014

Abstract

Saccharomyces cerevisiae shows a Crabtree effect that produces ethanol in a high glucose concentration even under fully aerobic condition. For efficient production of cake yeast or compressed yeast for baking, ethanol by-production is not desired since glucose limited chemostat or fed-batch cultivations are performed to suppress the Crabtree effect. In this study, the (13)C-based metabolic flux analysis ((13)C-MFA) was performed for the S288C derived S. cerevisiae strain to characterize a metabolic state under the reduced Crabtree effect. S. cerevisiae cells were cultured at a low dilution rate (0.1 h(-1)) under the glucose-limited chemostat condition. The estimated metabolic flux distribution showed that the acetyl-CoA in mitochondria was mainly produced from pyruvate by pyruvate dehydrogenase (PDH) reaction and that the level of the metabolic flux through the pentose phosphate pathway was much higher than that of the Embden-Meyerhof-Parnas pathway, which contributes to high biomass yield at low dilution rate by supplying NADPH required for cell growth.

Keywords: (13)C-based metabolic flux analysis; Central carbon metabolism; Crabtree effect; Redox balance; Saccharomyces cerevisiae.

MeSH terms

Acetyl Coenzyme A / metabolism
Aerobiosis
Biomass
Ethanol / metabolism
Glucose / metabolism
Glycolysis
Metabolic Flux Analysis*
NADP / metabolism
Oxidation-Reduction
Pentose Phosphate Pathway
Pyruvate Dehydrogenase Complex / metabolism
Pyruvic Acid / metabolism
Saccharomyces cerevisiae / cytology
Saccharomyces cerevisiae / growth & development
Saccharomyces cerevisiae / metabolism*

Substances

Pyruvate Dehydrogenase Complex
Ethanol
NADP
Acetyl Coenzyme A
Pyruvic Acid
Glucose