title: Production of 4-hydroxybutyrate from succinate semialdehyde in butyrate biosynthesis in Porphyromonas gingivalis.
abstract: BACKGROUND
  Despite evidence demonstrating the importance of butyrate-producing bacteria in host health and disease, the characterization of enzymes responsible for butyrate production has not been fully elucidated in the periodontopathogen, Porphyromonas gingivalis.
  
  
  METHODS
  LC-MS/MS and colorimetric analyses were employed to enzymatically characterize recombinant PGN_0724 in P. gingivalis as a succinate semialdehyde reductase. The concentration of short chain fatty acids in the culture supernatant of the wild-type bacteria and a mutant strain lacking the PGN_0724 gene were quantified using GC-MS.
  
  
  RESULTS
  Incubation of recombinant PGN_0724 with succinate semialdehyde and NADH resulted in the production of 4-hydroxybutyrate as well as consumption of succinate semialdehyde. Double reciprocal plots showed that the reaction catalyzed by the PGN_0724 protein was associated with a ternary complex mechanism. The growth speed and final turbidity of the mutant strain were much lower than those of the wild-type cells. The capacity of the mutant strain to produce butyrate, isobutyrate, and isovalerate was 30%, 15%, and 45%, respectively, of that of the wild-type strain, while the mutant strain produced approximately 3.9-fold more propionate than the wild type.
  
  
  CONCLUSIONS
  The pathway responsible for butyrate production is important for the growth of P. gingivalis and appears to be associated with production of the other short chain fatty acids.
  
  
  CONCLUSIONS
  The aim of this study was to delineate the mechanisms involved in the production of 4-hydroxybutyrate, which is an intermediate in the biosynthetic pathway for production of butyrate, which is a virulence factor in P. gingivalis.
authors.affiliation: Department of Microbiology, School of Dentistry, Aichi Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya 464-8650, Japan. Electronic address: yasuoy@dpc.agu.ac.jp.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Yoshida
authors.affiliation: Department of Microbiology, School of Dentistry, Aichi Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya 464-8650, Japan; Department of Removable Prosthodontics, School of Dentistry, Aichi Gakuin University, 2-11 Suemori-dori, Chikusa-ku, Nagoya 454-8651, Japan.
authors.forename: Mitsunari
authors.initials: M
authors.lastname: Sato
authors.affiliation: Department of Microbiology, School of Dentistry, Aichi Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya 464-8650, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Nagano
authors.affiliation: Department of Microbiology, School of Dentistry, Aichi Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya 464-8650, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Hasegawa
authors.affiliation: Department of Molecular and Cellular Biology, 1 Kawazumi, Mizuho-ku, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Okamoto
authors.affiliation: Department of Microbiology, School of Dentistry, Aichi Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya 464-8650, Japan.
authors.forename: Fuminobu
authors.initials: F
authors.lastname: Yoshimura
mesh_terms: D000447:Aldehydes / Q000378:metabolism*; D002850:Chromatography, Gel; D006885:Hydroxybutyrates / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D000447:Aldehydes; D006885:Hydroxybutyrates; C111420:4-hydroxybutyric acid
keywords: 4-hydroxybutyrate; Butyrate; Porphyromonas gingivalis; Short chain fatty acid; Succinate semialdehyde