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. 1992 Mar;58(3):916–919. doi: 10.1128/aem.58.3.916-919.1992

Purification and characterization of an intracellular peroxidase from Streptomyces cyaneus.

A Mliki 1, W Zimmermann 1
PMCID: PMC195355  PMID: 1315499

Abstract

An intracellular peroxidase (EC 1.11.1.7) from Streptomyces cyaneus was purified to homogeneity. The enzyme had a molecular weight of 185,000 and was composed of two subunits of equal size. It had an isoelectric point of 6.1. The enzyme had a peroxidase activity toward o-dianisidine with a Km of 17.8 microM and a pH optimum of 5.0. It also showed catalase activity with a Km of 2.07 mM H2O2 and a pH optimum of 8.0. The purified enzyme did not catalyze C alpha-C beta bond cleavage of 1,3-dihydroxy-2-(2-methoxyphenoxy)-1-(4-ethoxy-3-methoxyphenyl) propane, a nonphenolic dimeric lignin model compound. The spectrum of the peroxidase showed a soret band at 405 nm, which disappeared after reduction with sodium dithionite, indicating that the enzyme is a hemoprotein. Testing the effects of various inhibitors on the enzyme activity showed that it is a bifunctional enzyme having catalase and peroxidase activities.

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Selected References

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  1. Adhi T. P., Korus R. A., Crawford D. L. Production of Major Extracellular Enzymes during Lignocellulose Degradation by Two Streptomycetes in Agitated Submerged Culture. Appl Environ Microbiol. 1989 May;55(5):1165–1168. doi: 10.1128/aem.55.5.1165-1168.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Claiborne A., Fridovich I. Purification of the o-dianisidine peroxidase from Escherichia coli B. Physicochemical characterization and analysis of its dual catalatic and peroxidatic activities. J Biol Chem. 1979 May 25;254(10):4245–4252. [PubMed] [Google Scholar]
  4. Garfin D. E. Isoelectric focusing. Methods Enzymol. 1990;182:459–477. doi: 10.1016/0076-6879(90)82037-3. [DOI] [PubMed] [Google Scholar]
  5. Goldstein D. B. A method for assay of catalase with the oxygen cathode. Anal Biochem. 1968 Sep;24(3):431–437. doi: 10.1016/0003-2697(68)90148-6. [DOI] [PubMed] [Google Scholar]
  6. Hildebraunt A. G., Roots I. Reduced nicotinamide adenine dinucleotide phosphate (NADPH)-dependent formation and breakdown of hydrogen peroxide during mixed function oxidation reactions in liver microsomes. Arch Biochem Biophys. 1975 Dec;171(2):385–397. doi: 10.1016/0003-9861(75)90047-8. [DOI] [PubMed] [Google Scholar]
  7. Hochman A., Shemesh A. Purification and characterization of a catalase-peroxidase from the photosynthetic bacterium Rhodopseudomonas capsulata. J Biol Chem. 1987 May 15;262(14):6871–6876. [PubMed] [Google Scholar]
  8. Knoch M., van Pée K. H., Vining L. C., Lingens F. Purification, properties and immunological detection of a bromoperoxidase-catalase from Streptomyces venezuelae and from a chloramphenicol-nonproducing mutant. J Gen Microbiol. 1989 Sep;135(9):2493–2502. doi: 10.1099/00221287-135-9-2493. [DOI] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Loprasert S., Negoro S., Okada H. Thermostable peroxidase from Bacillus stearothermophilus. J Gen Microbiol. 1988 Jul;134(7):1971–1976. doi: 10.1099/00221287-134-7-1971. [DOI] [PubMed] [Google Scholar]
  11. MARGOLIASH E., NOVOGRODSKY A., SCHEJTER A. Irreversible reaction of 3-amino-1:2:4-triazole and related inhibitors with the protein of catalase. Biochem J. 1960 Feb;74:339–348. doi: 10.1042/bj0740339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pasti M. B., Pometto A. L., 3rd, Nuti M. P., Crawford D. L. Lignin-solubilizing ability of actinomycetes isolated from termite (Termitidae) gut. Appl Environ Microbiol. 1990 Jul;56(7):2213–2218. doi: 10.1128/aem.56.7.2213-2218.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ramachandra M., Crawford D. L., Hertel G. Characterization of an extracellular lignin peroxidase of the lignocellulolytic actinomycete Streptomyces viridosporus. Appl Environ Microbiol. 1988 Dec;54(12):3057–3063. doi: 10.1128/aem.54.12.3057-3063.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Takeda A., Miyahara T., Hachimori A., Samejima T. The interactions of thiol compounds with porcine erythrocyte catalase. J Biochem. 1980 Feb;87(2):429–439. doi: 10.1093/oxfordjournals.jbchem.a132763. [DOI] [PubMed] [Google Scholar]
  15. Wayne L. G., Diaz G. A. A double staining method for differentiating between two classes of mycobacterial catalase in polyacrylamide electrophoresis gels. Anal Biochem. 1986 Aug 15;157(1):89–92. doi: 10.1016/0003-2697(86)90200-9. [DOI] [PubMed] [Google Scholar]
  16. Zeiner R., Van Pée K. H., Lingens F. Purification and partial characterization of multiple bromoperoxidases from Streptomyces griseus. J Gen Microbiol. 1988 Dec;134(12):3141–3149. doi: 10.1099/00221287-134-12-3141. [DOI] [PubMed] [Google Scholar]
  17. van Pée K. H., Lingens F. Purification and molecular and catalytic properties of bromoperoxidase from Streptomyces phaeochromogenes. J Gen Microbiol. 1985 Aug;131(8):1911–1916. doi: 10.1099/00221287-131-8-1911. [DOI] [PubMed] [Google Scholar]

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