(13)C-metabolic flux analysis in S-adenosyl-L-methionine production by Saccharomyces cerevisiae.
S-Adenosyl-L-methionine (SAM) is a major biological methyl group donor, and is used as a nutritional supplement and prescription drug. Yeast is used for the industrial production of SAM owing to its high intracellular SAM concentrations. To determine the regulation mechanisms responsible for such high SAM production, (13)C-metabolic flux analysis ((13)C-MFA) was conducted to compare the flux distributions in the central metabolism between Kyokai no. 6 (high SAM-producing) and S288C (control) strains. (13)C-MFA showed that the levels of tricarboxylic acid (TCA) cycle flux in SAM-overproducing strain were considerably increased compared to those in the S228C strain. Analysis of ATP balance also showed that a larger amount of excess ATP was produced in the Kyokai 6 strain because of increased oxidative phosphorylation. These results suggest that high SAM production in Kyokai 6 strains could be attributed to enhanced ATP regeneration with high TCA cycle fluxes and respiration activity. Thus, maintaining high respiration efficiency during cultivation is important for improving SAM production.
Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan; 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.
Kenshi
K
Hayakawa
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.
Shuichi
S
Kajihata
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.
Fumio
F
Matsuda
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.
Hiroshi
H
Shimizu
D000255:Adenosine Triphosphate / Q000378:metabolism; D019069:Cell Respiration; D002952:Citric Acid Cycle; D064688:Metabolic Flux Analysis; D010085:Oxidative Phosphorylation; D012436:S-Adenosylmethionine / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000145:classification / Q000378:metabolism*
D012436:S-Adenosylmethionine; D000255:Adenosine Triphosphate
(13)C-based metabolic flux analysis; Central carbon metabolism; Crabtree effect; Redox balance; Saccharomyces cerevisiae