Applied Microbiology and Biotechnology

, Volume 40, Issue 6, pp 867–872 | Cite as

Biosynthesis of poly(γ-glutamic acid) from l-glutamic acid, citric acid, and ammonium sulfate in Bacillus subtilis IFO3335

  • Masao Kunioka
  • Atsuo Goto
Applied Microbial and Cell Physiology

Abstract

Poly(γ-glutamic acid) (PGA) production in Bacillus subtilis IFO3335 was studied. When l-glutamic acid, citric acid, and ammonium sulfate were used as carbon and nitrogen sources, a large amount of PGA without a by-product such as a polysaccharide was produced. The time courses of cell growth, PGA, glutamic acid, and citric acid concentrations during cultivation were investigated. It was found that glutamic acid added to the medium was apparently not assimilated. It can be presumed that the glutamic acid unit in PGA is mainly produced from citric acid and ammonium sulfate. The PGA productivity was investigated at various concentrations of ammonium sulfate in the media, which caused the depression of cell growth, high productivity of PGA, and the production of PGA with a high relative molecular mass. The yield of PGA determined by gel permeation chromatography (GPC) reached approximately 20 g/l. This yield was the highest value for PGA production by B. subtilis IFO3335, suggesting that B. subtilis IFO3335 was a bacterium that could produce PGA effectively. Time courses relative to the molecular mass of PGA at various concentrations of ammonium sulfate were investigated. It was suggested that B. subtilis IFO3335 excreted a PGA degradation enzyme with the progress of cultivation and that PGA was degraded by this enzyme.

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References

  1. Bovarnick M (1942) The formation of extracellular d(−)glutamic acid polypeptide by Bacillus subtilis. J Biol Chem 145:415–424Google Scholar
  2. Cheng C, Asada Y, Aida T (1989) Production of γ-polyglutamic acid by Bacillus subtilis A35 under denitrifying conditions. Agric Biol Chem 53:2369–2375Google Scholar
  3. Fujii H (1963) On the formation of mucilage by Bacillus natto. Part III Chemical constitutions of mucilage in natto (1). Nippon Nogeikagaku Kaishi 37:407–411Google Scholar
  4. Goto A, Kunioka M (1992) Biosynthesis and hydrolysis of poly(γ-glutamic acid) from Bacillus subtilis IFO3335. Biosci Biotech Biochem 56:1031–1035Google Scholar
  5. Hara T, Ueda S (1982) Regulation of polyglutamate production in Bacillus subtilis (natto); transformation of high PGA productivity. Agric Biol Chem 46:2275–2281Google Scholar
  6. Hara T, Aumayr A, Fujio Y, Ueda S (1982) Elimination of plasmid-linked polyglutamate production by Bacillus subtilis (natto) with acridine orange. Appl Environ Microbiol 44:1456–1458Google Scholar
  7. Ivánovics G, Bruckner V (1937) Chemische und immunologische Studien über den Mechanismus der Milzbrandinfektion und Immunität; die chemische Struktur der Kapselsubstanz des Milzbrandbazillus und der serologisch identischen spezifischen Substanz des Bacillus mesentericus. Z Immunitätsforsch 90:304–318Google Scholar
  8. Ivánovics G, Erdös L (1937) Ein Beitrag zum Wesen der Kapselsubstanz des Milzbrandbazillus. Z Immunitätsforsch 90:5–19Google Scholar
  9. Murao S (1969) On the polyglutamic acid fermentation. Kobunshi 16:1204–1212Google Scholar
  10. Murao S, Sawa S, Murakawa T, Omata S (1971) Polyglutamic acid fermentation part II. Culture conditions for the production of polyglutamic acid by Bacillus subtilis no. 5E. (1) Effect of amino acids and glucose. Nippon Nogeikagaku Kaishi 45:118–123Google Scholar
  11. Sawa S, Murao S, Murakawa T Omata S (1971) Polyglutamic acid fermentation part III. Culture conditions for the production of polyglutamic acid by Bacillus subtilis no. 5E. (2) Investigations on the synthetic media. Nippon Nogeikagaku Kaishi 45:124–129Google Scholar
  12. Thorne CB, Molnar DM (1955) d-Amino acid transamination in Bacillus anthracis. J Bacteriol 70:420–426Google Scholar
  13. Thorne CB, Gómez CG, Blind GR, Housewright RD (1953) Synthesis of glutamic acid and glutamyl polypeptide by Bacillus anthracis III. Factors affecting peptide production in synthetic liquid media. J Bacteriol 65:472–478Google Scholar
  14. Thorne CB, Gómez CG, Noyes HE, Housewright RD (1954) Production of glutamyl polypeptide by Bacillus subtilis. J Bacteriol 68:307–315Google Scholar
  15. Troy FA (1973) Chemistry and biosynthesis of the poly(γ-d-glutamyl) capsule in Bacillus subtilis. 1. Properties of the membrane-mediated biosynthetic reaction. J Biol Chem 248:305–315Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Masao Kunioka
    • 1
  • Atsuo Goto
    • 2
  1. 1.National Institute of Materials and Chemical ResearchTsukuba-shi, IbarakiJapan
  2. 2.Mitsubishi Paper Mills LimitedTsukuba Research LaboratoriesTsukuba-shi, IbarakiJapan

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