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J Biosci Bioeng. 2016 Apr;121(4):413-9. doi: 10.1016/j.jbiosc.2015.08.012. Epub 2015 Sep 26.

Metabolome analysis reveals the effect of carbon catabolite control on the poly(γ-glutamic acid) biosynthesis of Bacillus licheniformis ATCC 9945.

Author information

1
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.
2
AVT - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, 52074 Aachen, Germany. Electronic address: Lena.Meissner@avt.rwth-aachen.de.
3
AVT - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, 52074 Aachen, Germany. Electronic address: Thomas.Palmen@avt.rwth-aachen.de.
4
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.
5
AVT - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, 52074 Aachen, Germany. Electronic address: Jochen.Buechs@avt.rwth-aachen.de.
6
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.

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.

KEYWORDS:

Bacillus licheniformis ATCC 9945; Carbon catabolite control; Ion-pair-liquid chromatography coupled with tandem mass spectrometry; Metabolomics; Poly(γ-glutamic acid)

PMID:
26419706
DOI:
10.1016/j.jbiosc.2015.08.012
[Indexed for MEDLINE]

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