title: Metabolome analysis reveals the effect of carbon catabolite control on the poly(γ-glutamic acid) biosynthesis of Bacillus licheniformis ATCC 9945.
abstract: Poly(γ-glutamic acid) (PGA) is a polymer composed of L- and/or D-glutamic acids that is produced by Bacillus sp. Because the polymer has various features as water soluble, edible, non-toxic and so on, it has attracted attention as a candidate for many applications such as foods, cosmetics and so on. However, although it is well known that the intracellular metabolism of Bacillus sp. is mainly regulated by catabolite control, the effect of the catabolite control on the PGA producing Bacillus sp. is largely unknown. This study is the first report of metabolome analysis on the PGA producing Bacillus sp. that reveals the effect of carbon catabolite control on the metabolism of PGA producing Bacillus licheniformis ATCC 9945. Results showed that the cells cultivated in glycerol-containing medium showed higher PGA production than the cells in glucose-containing medium. Furthermore, metabolome analysis revealed that the activators of CcpA and CodY, global regulatory proteins of the intracellular metabolism, accumulated in the cells cultivated in glycerol-containing and glucose-containing medium, respectively, with CodY apparently inhibiting PGA production. Moreover, the cells seemed to produce glutamate from citrate and ammonium using glutamine synthetase/glutamate synthase. Pulsed addition of di-ammonium hydrogen citrate, as suggested by the metabolome result, was able to achieve the highest value so far for PGA production in B. licheniformis.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871 Osaka, Japan. Electronic address: hitoshi_mitsunaga@bio.eng.osaka-u.ac.jp.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Mitsunaga
authors.affiliation: AVT - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, 52074 Aachen, Germany. Electronic address: Lena.Meissner@avt.rwth-aachen.de.
authors.forename: Lena
authors.initials: L
authors.lastname: Meissner
authors.affiliation: AVT - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, 52074 Aachen, Germany. Electronic address: Thomas.Palmen@avt.rwth-aachen.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Palmen
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871 Osaka, Japan; Research Center for Transomics Medicine, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8285 Fukuoka, Japan. Electronic address: bamba@bio.eng.osaka-u.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Bamba
authors.affiliation: AVT - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, 52074 Aachen, Germany. Electronic address: Jochen.Buechs@avt.rwth-aachen.de.
authors.forename: Jochen
authors.initials: J
authors.lastname: Büchs
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, 565-0871 Osaka, Japan. Electronic address: fukusaki@bio.eng.osaka-u.ac.jp.
authors.forename: Eiichiro
authors.initials: E
authors.lastname: Fukusaki
mesh_terms: D064751:Ammonium Compounds / Q000378:metabolism; D001407:Bacillus / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D002244:Carbon / Q000378:metabolism*; D019343:Citric Acid / Q000031:analogs & derivatives / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005970:Glutamate Synthase / Q000378:metabolism; D018698:Glutamic Acid / Q000096:biosynthesis; D005990:Glycerol / Q000378:metabolism; D055442:Metabolome / Q000502:physiology*; D055432:Metabolomics; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000096:biosynthesis; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D064751:Ammonium Compounds; D001426:Bacterial Proteins; D000644:Quaternary Ammonium Compounds; D014157:Transcription Factors; C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; D019343:Citric Acid; D018698:Glutamic Acid; D002244:Carbon; D005970:Glutamate Synthase; D005947:Glucose; C426729:diammonium hydrogen citrate; D005990:Glycerol
keywords: Bacillus licheniformis ATCC 9945; Carbon catabolite control; Ion-pair-liquid chromatography coupled with tandem mass spectrometry; Metabolomics; Poly(γ-glutamic acid)