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Volume 148, Issue 6

A novel extracytoplasmic phenol oxidase of : its possible involvement in the onset of morphogenesis

The DDBJ accession number for the sequence reported in this paper is AB056583.

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Abstract

Exogenous addition of copper stimulates cellular differentiation in spp. Several lines of evidence suggested a parallel correlation between the stimulatory effect of copper and phenol-oxidizing enzyme activities in . Here a novel extracytoplasmic phenol oxidase (EpoA) associated with cellular development of this organism was identified and characterized. EpoA activity, examined by an in-gel stain procedure with ,′-dimethyl--phenylenediamine sulfate as a substrate, was repressed by glucose and induced by copper supplied in the medium. The enzyme activity was abolished and markedly reduced in the mutants forA-factor biosynthesis and , respectively, which suggested that the activity of the enzyme depends on those essential regulators for morphogenesis in . EpoA protein was purified to homogeneity and the N-terminal amino acid sequence was determined. A homologous sequence identified in the genomic database of A3(2) was used as a probe to clone the complete gene of . The deduced amino acid sequence of EpoA revealed that the mature protein with a molecular mass of 34 kDa was preceded by a signal peptide consisting of 34 aa, consistent with EpoA being a secreted enzyme. EpoA was predicted to be a laccase-type oxidase by not only the sequence similarity, but its substrate selectivity, oxidizing not tyrosine but dihydroxyphenylalanine (DOPA) to generate melanin pigment. Introduction of on a plasmid partially restored both the EpoA activity and aerial mycelium productivity in an A-factor-deficient mutant. Exogenous supplementation of a substance synthesized by purified EpoA from DOPA stimulated cellular differentiation in and several other species. Ultrafiltration indicated that the molecular mass of the putative stimulant synthesized by EpoA is between 500 and 1000 Da.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): BCDA, bathocuproinedisulfonic acid , copper , DOPA, dihydroxyphenylalanine , EpoA , EpoA, extracytoplasmic phenol oxidase , laccase , melanin , morphological differentiation , PMP, p-methoxyphenol and Streptomyces griseus
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2002-06-01
2024-12-14
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References

  1. Alexandre G., Zhulin I. B. 2000; Laccases are widespread in bacteria. Trends Biotechnol 18:41–42 [CrossRef]
     [Google Scholar]
  2. Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. 1982; Nucleotide sequence and exact localisation of the neomycin phosphotransferase gene from transposon Tn 5 . Gene 19:327–336 [CrossRef]
     [Google Scholar]
  3. Bell A. A., Wheeler M. H. 1986; Biosynthesis and functions of fungal melanins. Annu Rev Phytopathol 24:411–451 [CrossRef]
     [Google Scholar]
  4. Bibb M. J., Findlay P. R., Johnson M. W. 1984; The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene 30:157–166 [CrossRef]
     [Google Scholar]
  5. Chater K. F. 1993; Genetics of differentiation in Streptomyces . Annu Rev Microbiol 47:685–713 [CrossRef]
     [Google Scholar]
  6. Edens W. A., Goins T. Q., Dooley D., Henson J. M. 1999; Purification and characterization of a secreted laccase of Gaeumannomyces graminis var. tritici . Appl Environ Microbiol 65:3071–3074
     [Google Scholar]
  7. Endo K., Kamo K., Hosono K., Beppu T., Ueda K. 2001; Characterization of mutants defective in melanogenesis and a gene for tyrosinase of Streptomyces griseus . J Antibiot 54:789–796 [CrossRef]
     [Google Scholar]
  8. Georgellis D., Kwon O., Lin C. C. 2001; Quinones as the redox signal for the Arc two-component system of bacteria. Science 292:2314–2316 [CrossRef]
     [Google Scholar]
  9. Hofmann K., Bucher P., Falquet L., Bairoch A. 1999; The prosite database, its status in 1999. Nucleic Acids Res 27:215–219 [CrossRef]
     [Google Scholar]
  10. Hong S.-K., Matsumoto A., Horinouchi S., Beppu T. 1993; Effects of protein kinase inhibitors on in vitro protein phosphorylation and cellular differentiation of Streptomyces griseus . Mol Gen Genet 236:347–354
     [Google Scholar]
  11. Hopwood D. A., Bibb M. J., Chater K. F. 7 other authors 1985 Genetic Manipulation of Streptomyces: a Laboratory Manual Norwich: John Innes Foundation;
     [Google Scholar]
  12. Horinouchi S. 1999; γ-Butyrolactones that control secondary metabolism and cell differentiation in Streptomyces . In Cell–Cell Signaling in Bacteria pp 193–207 Edited by Dunny G. M., Winans S. C. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  13. Horinouchi S., Kumada Y., Beppu T. 1984; Unstable genetic determinant of A-factor biosynthesis in streptomycin-producing organisms: cloning and characterization. J Bacteriol 158:481–487
     [Google Scholar]
  14. Hullo M. F., Moszer I., Danchin A., Martin-Verstraete I. 2001; CotA of Bacillus subtilis is a copper-dependent laccase. J Bacteriol 183:5426–5430 [CrossRef]
     [Google Scholar]
  15. Keijer B. J. F., Wezel G. P., Canters G. W., Kieser T., Vijgenboom E. 2001; The ram -dependence of Streptomyces lividans differentiation is bypassed by copper. J Mol Microbiol Biotechnol 2:565–574
     [Google Scholar]
  16. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  17. Matvienko M., Torres M. J., Yoder J. I. 2001; Transcriptional responses in the hemiparasitic plant Triphysaria versicolor to host plant signals. Plant Physiol 127:272–282 [CrossRef]
     [Google Scholar]
  18. Miyadoh S. 1993; Research on antibiotic screening in Japan over the last decade: a producing microorganisms approach. Actinomycetologica 7:100–106 [CrossRef]
     [Google Scholar]
  19. Palmieri G., Giardina P., Bianco C., Fontanella B., Sannia G. 2000; Copper induction of laccase isoenzymes in ligninolytic fungus Pleurotus ostreatus . Appl Environ Microbiol 66:920–924 [CrossRef]
     [Google Scholar]
  20. Solano F., Lucas-Elio P., Fernandez E., Sanchez-Amat A. 2000; Marinomonas mediterranea MMB-1 transposon mutagenesis: isolation of a mutipotent polyphenol oxidase mutant. J Bacteriol 182:3754–3760 [CrossRef]
     [Google Scholar]
  21. Thompson J. D., Higgins D. G., Gibson T. J. 1994; clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680 [CrossRef]
     [Google Scholar]
  22. Thurston C. F. 1994; The structure and function of fungal laccases. Microbiology 140:19–26 [CrossRef]
     [Google Scholar]
  23. Tomita K., Oda N., Ohbayashi M., Kamei H., Miyaki T., Oki T. 1990; A new screening method for melanin biosynthesis inhibitors using Streptomyces bikiniensis . J Antibiot 41:1601–1605
     [Google Scholar]
  24. Ueda K., Miyake K., Horinouchi S., Beppu T. 1993; A gene cluster involved in aerial mycelium formation in Streptomyces griseus encodes proteins similar to the response regulator and membrane translocator. J Bacteriol 175:2006–2016
     [Google Scholar]
  25. Ueda K., Tomaru Y., Endoh K., Beppu T. 1997; Stimulatory effect of copper on antibiotic production and morphological differentiation in Streptomyces tanashiensis . J Antibiot 50:693–695 [CrossRef]
     [Google Scholar]
  26. Ueda K., Hsheh C.-W., Tosaki T., Shinkawa H., Beppu T., Horinouchi S. 1998; Characterization of an A-factor-responsive repressor for amfR essential for onset of aerial mycelium formation in Streptomyces griseus . J Bacteriol 180:5085–5093
     [Google Scholar]
  27. Xudong Z., Gibbons J., Garcia-Rivena J., Casadevall A., Williamson P. R. 2001; Laccase of Cryptococcus neoformans is a cell wall-associated virulence factor. Infect Immun 69:5589–5596 [CrossRef]
     [Google Scholar]
  28. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119 [CrossRef]
     [Google Scholar]
  29. Zak B. 1958; Simple procedure for the single sample determination of serum copper and iron. Clin Chim Acta 3:328–334 [CrossRef]
     [Google Scholar]
/content/journal/micro/10.1099/00221287-148-6-1767
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A novel extracytoplasmic phenol oxidase of Streptomyces: its possible involvement in the onset of morphogenesisThe DDBJ accession number for the sequence reported in this paper is AB056583.
Microbiology 148, 1767 (2002); https://doi.org/10.1099/00221287-148-6-1767
/content/journal/micro/10.1099/00221287-148-6-1767
/content/journal/micro/10.1099/00221287-148-6-1767
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