Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Dec;169(12):5575–5578. doi: 10.1128/jb.169.12.5575-5578.1987

Actinomycin synthesis in Streptomyces antibioticus: enzymatic conversion of 3-hydroxyanthranilic acid to 4-methyl-3-hydroxyanthranilic acid.

G H Jones 1
PMCID: PMC213988  PMID: 2445729

Abstract

A methyltransferase which utilizes 3-hydroxyanthranilic acid (HAA) as a substrate was identified in detergent-treated extracts of the bacterium Streptomyces antibioticus. The enzyme catalyzes the transfer of methyl groups from [14C]S-adenosylmethionine to HAA, but does not catalyze the methylation of 3-hydroxy-DL-kynurenine. Enzyme, substrate, time, and pH dependencies for the methyl transfer reaction were examined. Reaction products obtained from scaled-up reaction mixtures were fractionated by chromatography on Dowex 1, and the Dowex 1 fractions were examined by paper and thin-layer chromatography. One Dowex fraction was shown to contain a radioactive product with the chromatographic properties of 4-methyl-3-hydroxyanthranilic acid (MHA), a known intermediate in the biosynthesis of actinomycin. Available evidence indicates that the conversion of HAA to MHA is an early step in the biosynthesis of actinomycin by S. antibioticus and other actinomycin-producing streptomycetes.

Full text

PDF
5575

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Brown D., Hitchcock M. J., Katz E. Purification and characterization of kynurenine formamidase activities from Streptomyces parvulus. Can J Microbiol. 1986 Jun;32(6):465–472. doi: 10.1139/m86-086. [DOI] [PubMed] [Google Scholar]
  3. Choy H. A., Jones G. H. Phenoxazinone synthase from Streptomyces antibiotics: purification of the large and small enzyme forms. Arch Biochem Biophys. 1981 Oct 1;211(1):55–65. doi: 10.1016/0003-9861(81)90429-x. [DOI] [PubMed] [Google Scholar]
  4. Gallo M., Katz E. Regulation of secondary metabolite biosynthesis: catabolite repression of phenoxazinone synthase and actinomycin formation by glucose. J Bacteriol. 1972 Feb;109(2):659–667. doi: 10.1128/jb.109.2.659-667.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jones G. H., Hopwood D. A. Activation of phenoxazinone synthase expression in Streptomyces lividans by cloned DNA sequences from Streptomyces antibioticus. J Biol Chem. 1984 Nov 25;259(22):14158–14164. [PubMed] [Google Scholar]
  6. Jones G. H., Hopwood D. A. Molecular cloning and expression of the phenoxazinone synthase gene from Streptomyces antibioticus. J Biol Chem. 1984 Nov 25;259(22):14151–14157. [PubMed] [Google Scholar]
  7. Jones G. H. Regulation of phenoxazinone synthase expression in Streptomyces antibioticus. J Bacteriol. 1985 Sep;163(3):1215–1221. doi: 10.1128/jb.163.3.1215-1221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. KATZ E., WEISSBACH H. Biosynthesis of the actinomycin chromophore; enzymatic conversion of 4-methyl-3-hydroxyanthranilic acid to actinocin. J Biol Chem. 1962 Mar;237:882–886. [PubMed] [Google Scholar]
  9. KATZ E., WEISSBACH H. Incorporation of C14-labeled amino acids into actinomycin and protein by Streptomyces antibioticus. J Biol Chem. 1963 Feb;238:666–675. [PubMed] [Google Scholar]
  10. Keller U. Actinomycin synthetases. Multifunctional enzymes responsible for the synthesis of the peptide chains of actinomycin. J Biol Chem. 1987 Apr 25;262(12):5852–5856. [PubMed] [Google Scholar]
  11. Keller U. Acyl pentapeptide lactone synthesis in actinomycin-producing streptomycetes by feeding with structural analogs of 4-methyl-3-hydroxyanthranilic acid. J Biol Chem. 1984 Jul 10;259(13):8226–8231. [PubMed] [Google Scholar]
  12. Keller U., Kleinkauf H., Zocher R. 4-Methyl-3-hydroxyanthranilic acid activating enzyme from actinomycin-producing Streptomyces chrysomallus. Biochemistry. 1984 Mar 27;23(7):1479–1484. doi: 10.1021/bi00302a022. [DOI] [PubMed] [Google Scholar]
  13. Mason K. T., Shaw G. J., Katz E. Biosynthetic studies with L-[2,3-3H2] valine as precursor of the D-valine moiety in actinomycin. Arch Biochem Biophys. 1977 Apr 30;180(2):509–513. doi: 10.1016/0003-9861(77)90066-2. [DOI] [PubMed] [Google Scholar]
  14. Perlman D., Otani S., Perlman K. L., Walker J. E. 3-Hydroxy-4-methylkynurenine as an intermediate in actinomycin biosynthesis. J Antibiot (Tokyo) 1973 May;26(5):289–296. doi: 10.7164/antibiotics.26.289. [DOI] [PubMed] [Google Scholar]
  15. SIVAK A., MELONI M. L., NOBILI F., KATZ E. Biosynthesis of the actinomycin chromophore. Studies with DL-[7alpha-14C] tryptophan and L-[Me-14C] methionine. Biochim Biophys Acta. 1962 Feb 26;57:283–289. doi: 10.1016/0006-3002(62)91121-6. [DOI] [PubMed] [Google Scholar]
  16. Troost T., Hitchcock M. J., Katz E. Distinct kynureninase and hydroxykynureninase enzymes in an actinomycin-producing strain of Streptomyces parvulus. Biochim Biophys Acta. 1980 Mar 14;612(1):97–106. doi: 10.1016/0005-2744(80)90282-x. [DOI] [PubMed] [Google Scholar]
  17. Troost T., Katz E. Phenoxazinone biosynthesis: accumulation of a precursor, 4-methyl-3-hydroxyanthranilic acid, by mutants of Streptomyces parvulus. J Gen Microbiol. 1979 Mar;111(1):121–132. doi: 10.1099/00221287-111-1-121. [DOI] [PubMed] [Google Scholar]
  18. WEISSBACH H., REDFIELD B., BEAVEN V., KATZ E. 4-METHYL-3-HYDROXYANTHRANILIC ACID, AN INTERMEDIATE IN ACTINOMYCIN BIOSYNTHESIS. Biochem Biophys Res Commun. 1965 May 3;19:524–530. doi: 10.1016/0006-291x(65)90157-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES