title: Enhancement of 3-hydroxypropionic acid production from glycerol by using a metabolic toggle switch.
abstract: BACKGROUND
  3-hydroxypropionic acid (3-HP) is an important platform for the production of C3 chemicals, including acrylic acid, methyl acrylate, and acrylamide. Microbial production of 3-HP is mainly due to glycerol metabolism. In this study, in order to improve microbial 3-HP production, we applied a metabolic toggle switch for controlling the glycerol metabolism to redirect the excess metabolic flux of central metabolic pathway toward an exogenous 3-HP producing pathway in Escherichia coli.
  
  
  RESULTS
  The metabolic toggle switch enables conditional repression of the expression of a target gene during the fermentation. We individually performed conditional repression of glpK, tpiA, and gapA, which are involved in glycerol metabolism. The conditional repression of glpK and tpiA was not effective for 3-HP production under our experimental conditions. However, gapA conditional repression contributed to improve 3-HP production (titer, 54.2 ± 1.5 mM; yield, 32.1 ± 1.3 %) compared with that for the wild type strain. Additional deletion of endogenous yqhD, which is responsible for the production of a major byproduct, 1,3-propandiol, further increased 3-HP production (titer, 67.3 ± 2.1 mM; yield, 51.5 ± 3.2 %). The titer and yield were 80 and 94 % higher than those of the wild type strain, respectively. The obtained 3-HP yield from glycerol is comparable with the highest yield ever reported for microbial 3-HP production using glycerol as a sole carbon source. The measurement of intracellular metabolites showed the metabolic toggle switch successfully controlled the metabolic flux.
  
  
  CONCLUSIONS
  The conditional repression of gapA by using the metabolic toggle switch combined with deletion of endogeneous yqhD increased 3-HP production approximately twofold from glycerol. This result indicates the metabolic toggle switch can be applied in various bio-production using diverse substrates.
authors.affiliation: Laboratory for Bioinformatics, Graduate School of Systems Life Sciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. ktsuruno@brs.kyushu-u.ac.jp.
authors.forename: Keigo
authors.initials: K
authors.lastname: Tsuruno
authors.affiliation: Laboratory for Bioinformatics, Graduate School of Systems Life Sciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. honjo@brs.kyushu-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Honjo
authors.affiliation: Laboratory for Bioinformatics, Graduate School of Systems Life Sciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. taizo@brs.kyushu-u.ac.jp.
authors.forename: Taizo
authors.initials: T
authors.lastname: Hanai
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002952:Citric Acid Cycle; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism*; D006019:Glycolysis; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D013058:Mass Spectrometry; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways; D010957:Plasmids / Q000235:genetics / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D019344:Lactic Acid; C031601:hydracrylic acid; D005990:Glycerol