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Study of the role of anaerobic metabolism in succinate production by Enterobacter aerogenes

  • Biotechnological products and process engineering
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Abstract

Succinate is a core biochemical building block; optimizing succinate production from biomass by microbial fermentation is a focus of basic and applied biotechnology research. Lowering pH in anaerobic succinate fermentation culture is a cost-effective and environmentally friendly approach to reducing the use of sub-raw materials such as alkali, which are needed for neutralization. To evaluate the potential of bacteria-based succinate fermentation under weak acidic (pH <6.2) and anaerobic conditions, we characterized the anaerobic metabolism of Enterobacter aerogenes AJ110637, which rapidly assimilates glucose at pH 5.0. Based on the profile of anaerobic products, we constructed single-gene knockout mutants to eliminate the main anaerobic metabolic pathways involved in NADH re-oxidation. These single-gene knockout studies showed that the ethanol synthesis pathway serves as the dominant NADH re-oxidation pathway in this organism. To generate a metabolically engineered strain for succinate production, we eliminated ethanol formation and introduced a heterogeneous carboxylation enzyme, yielding E. aerogenes strain ΔadhE/PCK. The strain produced succinate from glucose with a 60.5 % yield (grams of succinate produced per gram of glucose consumed) at pH <6.2 and anaerobic conditions. Thus, we showed the potential of bacteria-based succinate fermentation under weak acidic conditions.

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Acknowledgments

We thank Yoshihiko Hara and Dr. Akito Chinen for providing experimental materials and Yoko Yamamoto for useful discussion. We also thank Techno Suruga Co., Ltd., for providing technical support for the phenotypic and genetic characterization of AJ110637. This study was funded by Ajinomoto Co. Inc., and no external funds were used.

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Authors and Affiliations

  1. Institute for Innovation, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan

    Yoshinori Tajima, Kenichi Kaida, Atsushi Hayakawa, Keita Fukui, Yousuke Nishio & Yoshihiro Usuda

  2. Research Institute for Bioscience Product & Fine Chemicals, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan

    Kenichi Hashiguchi

  3. Japan Bioindustry Association, 2-26-9 Hatchobori, Chuo-Ku, Tokyo, 104-0032, Japan

    Ryosuke Fudou

  4. Business Strategy and Development Department, Ajinomoto Co., Inc., 15-1 Kyobashi, 1-Chome, Chuo-Ku, Tokyo, 104-8315, Japan

    Kazuhiko Matsui

  5. Department of Biotechnology, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan

    Koji Sode

Authors
  1. Yoshinori Tajima
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  2. Kenichi Kaida
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  3. Atsushi Hayakawa
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  4. Keita Fukui
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  8. Kazuhiko Matsui
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  9. Yoshihiro Usuda
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Correspondence to Yoshinori Tajima.

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Tajima, Y., Kaida, K., Hayakawa, A. et al. Study of the role of anaerobic metabolism in succinate production by Enterobacter aerogenes . Appl Microbiol Biotechnol 98, 7803–7813 (2014). https://doi.org/10.1007/s00253-014-5884-3

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  • DOI: https://doi.org/10.1007/s00253-014-5884-3

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