TY - JOUR
T1 - Pyruvate metabolism redirection for biological production of commodity chemicals in aerobic fungus Aspergillus oryzae
AU - Zhang, Silai
AU - Wakai, Satoshi
AU - Sasakura, Naoya
AU - Tsutsumi, Hiroko
AU - Hata, Yoji
AU - Ogino, Chiaki
AU - Kondo, Akihiko
N1 - Funding Information:
The authors gratefully acknowledge Honorary Professor Katsuhiko Kitamoto (Tokyo University) for providing the A. oryzae strain NSPlD1 and Professor Katuya Gomi (Tohoku University) for providing the A. oryzae strain ΔligD::sC. This work was supported by Special Coordination Funds for Promoting Science and Technology, and by the Creation of Innovation Centers for Advanced Interdisciplinary Research Areas program (Innovative Bioproduction Kobe) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) , Japan.
Funding Information:
The authors gratefully acknowledge Honorary Professor Katsuhiko Kitamoto (Tokyo University) for providing the A. oryzae strain NSPlD1 and Professor Katuya Gomi (Tohoku University) for providing the A. oryzae strain ΔligD::sC. This work was supported by Special Coordination Funds for Promoting Science and Technology, and by the Creation of Innovation Centers for Advanced Interdisciplinary Research Areas program (Innovative Bioproduction Kobe) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), Japan.
Publisher Copyright:
© 2020 International Metabolic Engineering Society
PY - 2020/9
Y1 - 2020/9
N2 - Pyruvate is a central metabolite for the biological production of various chemicals. In eukaryotes, pyruvate produced by glycolysis is used in conversion to ethanol and lactate and in anabolic metabolism in the cytosol, or is transported into the mitochondria for use as a substrate in the tricarboxylic acid (TCA) cycle. In this study, we focused on controlling pyruvate metabolism in aerobic microorganisms for the biological production of various chemicals. We successfully improved productivity by redirecting pyruvate metabolism in the aerobic filamentous fungus Aspergillus oryzae via the deletion of two genes that encode pyruvate decarboxylase and mitochondrial pyruvate carriers. Production of ethanol as a major byproduct was completely inhibited, and the limited translocation of pyruvate into the mitochondria shifted the metabolism from respiration for energy conversion to the effective production of lactate or 2,3-butandiole, even under aerobic conditions. Metabolomic and transcriptomic analyses showed an emphasis on glycolysis and a repressed TCA cycle. Although the dry mycelial weights of the deletion mutants were reduced compared with those of wild type, the titer and yields of the target products were drastically increased. In particular, the redirection of pyruvate metabolism shifted from anabolism for biomass production to catabolism for the production of target chemicals. Conclusively, our results indicate that the redirection of pyruvate metabolism is a useful strategy in the metabolic engineering of aerobic microorganisms.
AB - Pyruvate is a central metabolite for the biological production of various chemicals. In eukaryotes, pyruvate produced by glycolysis is used in conversion to ethanol and lactate and in anabolic metabolism in the cytosol, or is transported into the mitochondria for use as a substrate in the tricarboxylic acid (TCA) cycle. In this study, we focused on controlling pyruvate metabolism in aerobic microorganisms for the biological production of various chemicals. We successfully improved productivity by redirecting pyruvate metabolism in the aerobic filamentous fungus Aspergillus oryzae via the deletion of two genes that encode pyruvate decarboxylase and mitochondrial pyruvate carriers. Production of ethanol as a major byproduct was completely inhibited, and the limited translocation of pyruvate into the mitochondria shifted the metabolism from respiration for energy conversion to the effective production of lactate or 2,3-butandiole, even under aerobic conditions. Metabolomic and transcriptomic analyses showed an emphasis on glycolysis and a repressed TCA cycle. Although the dry mycelial weights of the deletion mutants were reduced compared with those of wild type, the titer and yields of the target products were drastically increased. In particular, the redirection of pyruvate metabolism shifted from anabolism for biomass production to catabolism for the production of target chemicals. Conclusively, our results indicate that the redirection of pyruvate metabolism is a useful strategy in the metabolic engineering of aerobic microorganisms.
KW - 2,3-Butanediol
KW - Aspergillus oryzae
KW - L-lactic acid
KW - Mitochondrial pyruvate carrier
KW - Pyruvate decarboxylase
KW - Pyruvate flux
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U2 - 10.1016/j.ymben.2020.06.010
DO - 10.1016/j.ymben.2020.06.010
M3 - Article
C2 - 32623009
AN - SCOPUS:85087522738
VL - 61
SP - 225
EP - 237
JO - Metabolic Engineering
JF - Metabolic Engineering
SN - 1096-7176
ER -