1887

Microbiology Society logo

Volume 142, Issue 7

Trypsin-like protease of Streptomyces exfoliatus SMF13, a potential agent in mycelial differentiation Free

Abstract

SMF13 sequentially produced leupeptin, leupeptin-inactivating enzyme (LIE) and trypsin-like protease (TLP). TLP was produced upon exhaustion of glucose. Autolysis of mycelium was accompanied by an increase in TLP activity. However, in three mutants isolated from SMF13 after UV-mutagenesis, mycelium autolysis did not occur, and neither LIE nor TLP was produced, although leupeptin was produced. Production of both LIE and TLP was restored in a spontaneous Spo revertant of a mutant. In contrast, two mutants sequentially produced leupeptin, LIE and TLP. The molecular mass of TLP produced during morphological differentiation was estimated to be 31.8 kDa by SOS-PAGE. The N-terminal amino acid sequence was RVGGTxAAQGNFPFQQxLSM. TLP was competitively inhibited by leupeptin; the inhibition constant was 0.015 μM. TLP effectively hydrolysed the mycelial protein extract of SMF13, but the hydrolytic activity was inhibited by leupeptin. It was concluded that morphological differentiation and production of TLP are coordinately regulated, that TLP may function as an enzyme in the metabolism of mycelial proteins, and that the hydrolytic activity of TLP is regulated by autogenous leupeptin in SMF13.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): leupeptin , morphological differentiation , regulation , Streptomyces exfoliatus and trypsin-like protease
© Society for General Microbiology 1996
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-7-1797
1996-07-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/7/mic-142-7-1797.html?itemId=/content/journal/micro/10.1099/13500872-142-7-1797&mimeType=html&fmt=ahah

References

  1. Aoyagi T. 1989; Protease inhibitor and biological control. In Bioactive Metabolites from Microorganisms : Progress in Industrial Microbiology 27 pp. 403–418 Edited by Bushell M. E., Grafe U. New York: Elsevier;
     [Google Scholar]
  2. Aoyagi T., Miyata S., Nanbo M., Kojima F., Matsuzaki M., Ishizuka M., Takeuchi T., Umezawa H. 1969; Biological activities of leupeptins.. J Antibiot 22:558–568
     [Google Scholar]
  3. Babcock M.J., Kendrick K. E. 1988; Cloning of DNA involved in sporulation of Streptomyces griseus. J Bacteriol 170:2802–2808
     [Google Scholar]
  4. Bascaran V., Hardisson C., Brana A. F. 1990; Regulation of extracellular protease production in Streptomyces clavuligerus.. Appl Microbiol biotechnol 34:208–213
     [Google Scholar]
  5. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
     [Google Scholar]
  6. Champness W.C. 1988; New loci required for Streptomyces coelicolor morphological and physiological differentiation. J Bacteriol 170:1168–1174
     [Google Scholar]
  7. Chandrasekaran S., Dhar S. C. 1987; Multiple proteases from Streptomyces moderatus. II. Isolation and purification of five extracellular proteases. Arch Biochem Biophys 257:402–408
     [Google Scholar]
  8. Chater K.F. 1984; Morphological and physiological differentiation in Streptomyces.. In Microbial Development pp. 89–115 Edited by Losick R., Shapiro L. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  9. Chauvet J., Dostal I., Acher R. 1976; Isolation of a trypsin-like enzyme from Streptomycesparomomycinus (paromotrypsin) by affinity adsorption through kunitz inhibitor-sepharose.. Int J Pept Protein Res 8:45–55
     [Google Scholar]
  10. Daza A., Martin J. F., Gil J. A. 1990; High transformation frequency of nonsporulating mutants of Streptomyces griseus. FEMS Microbiol Eett 71:259–264
     [Google Scholar]
  11. Gibb G.D., Strahl W. R. 1988; Physiological regulation of protease activity in Streptomyces peucetius. Can J Microbiol 34:187–190
     [Google Scholar]
  12. Ginther C.L. 1979; Sporulation and the production of serine protease and cephamycin C by Streptomyces lactamdurans. Antimicrob Agents Chemother 15:522–526
     [Google Scholar]
  13. Hatanaka Y., Tsunematsu H., Mizusaki K., Makisumi S. 1985; Interactions of derivatives of guanidinophenylalanine and guanidinophenylglycine with Streptomyces griseus trypsin. biochim biophys Acta 832:274–279
     [Google Scholar]
  14. Hopwood D. A., Bibb M. J., Chater K. F., Kieser T., Bruton C. J., Kieser H. M., Lydiate D. J., Smith C. P., Ward J. M., Schrempf H. 1985; Mutagenesis of Streptomyces spores by ultraviolet light. In Genetic Manipulation of Streptomyces: a Laboratory Manual pp. 37–38 Norwich: John Innes Foundation;
     [Google Scholar]
  15. Keil B. 1971; Trypsin. In The Enzymes 3 pp. 249–275 Edited by Boyer P. D. New York, London: Academic Press;
     [Google Scholar]
  16. Kim I.S., Lee K. J. 1995; Physiological roles of leupeptin and extracellular proteases in mycelium development of Streptomyces exfoliatus SMF13. Microbiology 141:1017–1025
     [Google Scholar]
  17. Kim I. S., Han Y. T., Barrow K. D., Lee K. J. 1993; The structure of protease inhibitors produced by Streptomyces exfoliatus SMF13.. Korean J Microbiol 31:326–334
     [Google Scholar]
  18. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
     [Google Scholar]
  19. Martin J.F., Demain A. L. 1980; Control of antibiotic biosynthesis. Microbiol Rev 44:230–251
     [Google Scholar]
  20. Matsudaira P. 1987; Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem 262:10035–10038
     [Google Scholar]
  21. Mendez C., Brana A. F., Manzanal M. B., Hardisson C. 1985; Role of substrate mycelium in colony development in Streptomyces. Can J Microbiol 31:446–450
     [Google Scholar]
  22. Merrick M.J. 1976; A morphological and genetic mapping study of bald colony mutants of Streptomyces coelicolor. J Gen Microbiol 96:299–315
     [Google Scholar]
  23. Miguelez E. M., Garcia M., Hardisson C., Manzanal M. B. 1994; Autoradiographic study of hyphal growth during aerial mycelium formation in Streptomyces antibioticus. J Bacteriol 176:2105–2107
     [Google Scholar]
  24. Miller G.L. 1959; Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
     [Google Scholar]
  25. Millónig G. 1961; Advantages of a phosphate of Os04 solutions in fixation.. J Appl Physiol 32:1637–1640
     [Google Scholar]
  26. Morihara K., Tsuzuki H. 1968; A trypsin-like protease from Streptomyces fradiae.. Arch biochem biophys 126:971–973
     [Google Scholar]
  27. Narahashi Y. 1970; Pronase. Methods Ensçymol 19:651–664
     [Google Scholar]
  28. Olafson R. W., Jurasek L., Carpenter M. R., Smillie L. B. 1975; Amino acid sequence of Streptomyces griseus trypsin. Cyanogen bromide fragments and complete sequence. Biochemistry 14:1168–1176
     [Google Scholar]
  29. Palubinskas V. I., Yankevich N. B., Yanulaitene K. K., Vesa V. S., Bendikene V. G., Maksimenko A.V., Torchilin M.P., Ilyina E. V., Smirnov V. N., Krestyanoe L. N., Bartoshevich Y. E., Zabirova R. C. 1984; Trypsin-like enzyme from Streptomyces 771. Purification and properties of native and immobilized enzyme.. Appl biochem biotechnol 9:231–241
     [Google Scholar]
  30. Peczynska-Czoch W., 81 Mordarski W. 1988; Actinomycete enzymes. In Actinomycetes in biotechnology pp. 246–250 Edited by Goodfellow M., Williams S. T. , Mordarski M. London: Academic Press;
     [Google Scholar]
  31. Pirt S.J. 1975; Parameters of growth and analysis of growth data. In Principles of Microbe and Cell Cultivation pp. 4–14 Oxford: Blackwell;
     [Google Scholar]
  32. Renko M., Vitale L. J., Kokalj M., Pokorny M. 1989; Streptomyces rimosus extracellular proteases. 4. Trypsin-like proteinase.. Appl Microbiol biotechnol 31:38–44
     [Google Scholar]
  33. Sarath G., De La Motte R.S., Wagner F. W. 1989; Protease assay methods. In Proteolytic Enzymes: a Practical Approach pp. 25–55 Edited by Beynone R. J., Bond J. S. Oxford: IRL Press;
     [Google Scholar]
  34. Wildermath H. 1970; Development and organization of the aerial mycelium in Streptomyces coelicolor. J Gen Microbiol 60:43–50
     [Google Scholar]
  35. Yoshida N., Sasaki A., Inoue H. 1971; An anionic trypsin-like enzyme from Streptomyces erythraeus.. FEbS Lett 15:129–132
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-7-1797
Loading
Trypsin-like protease of Streptomyces exfoliatus SMF13, a potential agent in mycelial differentiation
Microbiology 142, 1797 (1996); https://doi.org/10.1099/13500872-142-7-1797
/content/journal/micro/10.1099/13500872-142-7-1797
/content/journal/micro/10.1099/13500872-142-7-1797
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error