title: Impact of an energy-conserving strategy on succinate production under weak acidic and anaerobic conditions in Enterobacter aerogenes.
abstract: BACKGROUND
  Succinate is an important C4 building block chemical, and its production via fermentative processes in bacteria has many practical applications in the biotechnology field. One of the major goals of optimizing the bacterium-based succinate production process is to lower the culture pH from the current neutral conditions, as this would reduce total production costs. In our previous studies, we selected Enterobacter aerogenes, a rapid glucose assimilator at pH 5.0, in order to construct a metabolically engineered strain that could produce succinate under weakly acidic conditions. This engineered strain produced succinate from glucose with a 72.7% (g/g) yield at pH 5.7, with a volumetric productivity of 0.23 g/L/h. Although this demonstrates proof-of-concept that bacterium-based succinate fermentation can be improved under weakly acidic conditions, several parameters still required further optimization.
  
  
  RESULTS
  In this study, we genetically modified an E. aerogenes strain previously developed in our laboratory in order to increase the production of ATP during succinate synthesis, as we inferred that this would positively impact succinate biosynthesis. This led to the development of the ES08ΔptsG strain, which contains the following modifications: chromosomally expressed Actinobacillus succinogenes phosphoenolpyruvate carboxykinase, enhanced fumarate reductase, inactivated pyruvate formate lyase, pyruvate oxidase, and glucose-phosphotransferase permease (enzyme IIBC(Glc)). This strain produced 55.4 g/L succinate from glucose, with 1.8 g/L acetate as the major byproduct at pH 5.7 and anaerobic conditions. The succinate yield and volumetric productivity of this strain were 86.8% and 0.92 g/L/h, respectively.
  
  
  CONCLUSIONS
  Focusing on increasing net ATP production during succinate synthesis leads to increased succinate yield and volumetric productivity in E. aerogenes. We propose that the metabolically engineered E. aerogenes ES08ΔptsG strain, which effectively produces succinate under weakly acidic and anaerobic conditions, has potential utility for economical succinate production.
authors.affiliation: Institute for Innovation, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan. yoshinori_tajima@ajinomoto.com.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Tajima
authors.affiliation: Institute for Innovation, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan. youkoa_yamamoto@ajinomoto.com.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Yamamoto
authors.affiliation: Institute for Innovation, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan. keita_fukui@ajinomoto.com.
authors.forename: Keita
authors.initials: K
authors.lastname: Fukui
authors.affiliation: Institute for Innovation, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan. yousuke_nishio@ajinomoto.com.
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Nishio
authors.affiliation: Research Institute for Bioscience Product & Fine Chemicals, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, Japan. kenichi_hashiguchi@ajinomoto.com.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Hashiguchi
authors.affiliation: Institute for Innovation, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan. yoshihiro_usuda@ajinomoto.com.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Usuda
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Tokyo University of Agriculture and Technology, Naka-cho, Koganei, Tokyo, Japan. sode@cc.tuat.ac.jp.
authors.forename: Koji
authors.initials: K
authors.lastname: Sode
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000693:Anaerobiosis; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism; D021902:Enterobacter aerogenes / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D060847:Metabolic Engineering / Q000379:methods*; D019802:Succinic Acid / Q000378:metabolism*
chemical_list: D003470:Culture Media; D000255:Adenosine Triphosphate; D019802:Succinic Acid