Skip to main content
Log in

Isolation and characterization of an extracellular proteinase produced by a soil strain ofXanthomonas maltophilia

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

An extracellular proteinase from aXanthomonas maltophilia strain isolated from soil was purified by ammonium sulfate precipitation, gel exclusion, and ion exchange gel chromatography. The enzyme appeared homogeneous upon polyacrylamide-gel electrophoresis. Its molecular weight was estimated to be 36,000 by the gel filtration assay and 40,000 by polyacrylamidegel electrophoresis after denaturation: it was a monomeric protein. Highly active at alkaline pH with casein as the substrate, it was inactivated by serine-site inhibitors, such as phenylmethylsulfonyl fluoride and diisopropylfluorophosphate. Like serine proteases, it hydrolyzed esters such as the β-naphthyl acetate. Moreover, like metalloproteases, the enzyme was rapidly inactivated by ethylene diamine tetraacetate, and the activity was partially restored by calcium. These original characteristics have not often been reported among other gram-negative bacteria.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Literature Cited

  1. Andrews P (1965) The gel filtration behavior of proteins related to their molecular weight over a wide range. Biochem J 96:595–606

    PubMed  Google Scholar 

  2. Belew M, Porath J (1970) Extracellular proteinase fromPenicillium notatum. Methods Enzymol 19:576–581

    Google Scholar 

  3. Boethling RS (1975) Purification and properties of a serine protease fromPseudomonas maltophilia. J Bacteriol 121:933–941

    PubMed  Google Scholar 

  4. Bradford MM (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

    PubMed  Google Scholar 

  5. Braun V, Schmitz G (1980) Excretion of a protease bySerratia marcescens. Arch Microbial 24:55–61

    Article  Google Scholar 

  6. Breuil C, Kushner DJ (1975) Lipase and esterase formation by psychrophilic and mesophilicAcinetobacter species. Can J Microbiol 21:424–433

    Google Scholar 

  7. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann NY Acad Sci 121:404–427

    PubMed  Google Scholar 

  8. Debette J, Blondeau R (1977) Caractérisation de bactéries telluriques assimilables àPseudomonas maltophilia. Can J Microbiol 23:1123–1127

    PubMed  Google Scholar 

  9. Debette J, Blondeau R (1980) Présence dePseudomonas maltophilia dans la rhizosphère de quelques plantes cultivées. Can J Microbiol 26:460–463

    PubMed  Google Scholar 

  10. Debette J, Prensier G (1989) Immunoelectron microscopic demonstration of an esterase on the outer membrane ofXanthomonas maltophilia. Appl Environ Microbiol 55:233–239

    PubMed  Google Scholar 

  11. Debette J, Losfeld J, Blondeau R (1975) Taxonomie numérique de bactéries telluriques nonfermentantes à Gram-négatif. Can J Microbiol 21:1322–1334

    PubMed  Google Scholar 

  12. De Moreno MR, Smith JF, Smith RV (1985) Silver staining of proteins in polyacrylamide gels: increased sensitivity through a combined Coomassie blue-silver stain procedure. Anal Biochem 151:466–470

    Article  PubMed  Google Scholar 

  13. Denis F, Veillet-Poncet L (1984) Essai de mise en évidence d'une activité protéolytique d'origine intracellulaire chezAeromonas hydrophila LP 50. Can J Microbiol 30:1190–1192

    PubMed  Google Scholar 

  14. Fairbairn DJ, Law BA (1986) Proteinases of psychrotrophic bacteria: their production, properties, effects and control. J Dairy Res 53:139–177

    PubMed  Google Scholar 

  15. Gnosspelius G (1978) Purification and properties of an extracellular protease fromMyxococcus virescens. J Bacteriol 133:17–25

    PubMed  Google Scholar 

  16. Iizuka H, Komagata K (1964) Microbial studies on petroleum and natural gas. I. Determination of hydrocarbon utilizing bacteria. J Gen Appl Microbiol 10:207–221

    Google Scholar 

  17. Ikemoto S, Suzuki K, Kaneko T, Komagata K (1980) Characterization of strains ofPseudomonas maltophilia which do not require methionine. Int J Syst Bacteriol 30:437–447

    Google Scholar 

  18. Jensen SE, Phillippe L, Teng Tseng J, Stemke GW, Campbell JN (1980) Purification and characterization of exocellular proteases produced by a clinical isolate and a laboratory strain ofPseudomonas aeruginosa. Can J Microbiol 26:77–86

    PubMed  Google Scholar 

  19. Kato N, Nagasawa T, Adachi S, Tani Y, Ogata K (1972) Purification and properties of proteases from a marine-psychrophilic bacterium. Agr Biol Chem Tokyo 36:1185–1192

    Google Scholar 

  20. Kirkeby S, Moe D (1988) A comparison between activities for nonspecific esterases and ester proteases. Acta Histochem 83:11–20

    PubMed  Google Scholar 

  21. Kobayashi T, Ogasawara A, Ito S, Saitoh M (1985) Purification and some properties of alkaline proteinase produced byPseudomonas maltophilia. Agr Biol Chem Tokyo 49:693–698

    Google Scholar 

  22. Kraut J (1977) Serine proteases: structure and mechanism of catalysis. Annu Rev Biochem 46:331–358

    Article  PubMed  Google Scholar 

  23. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  Google Scholar 

  24. Lambert B, Leyns F, Van Rooyen L, Gosselé F, Papon Y, Swings J (1987) Rhizobacteria of maïze and their antifungal activities. Appl Environ Microbiol 53:1866–1871

    Google Scholar 

  25. Matsubara H, Feder J (1971) Other bacterial, mold and yeast proteases. In: Boyer P.D. (ed) The enzymes, vol 3, New York: Academic Press Inc pp 721–792

    Google Scholar 

  26. Moore WE, Cato EP, Moore LVH (1985) Index of the bacterial and yeast nomenclatural changes published in the International Journal of Systematic Bacteriology since the 1980 approved lists of bacterial names (1 January 1980 to 1 January 1985). Int J Syst Bacteriol 35:382–407

    Google Scholar 

  27. Morihara K (1974) Comparative specificity of microbial proteinases. Adv Enzymol 41:179–243

    PubMed  Google Scholar 

  28. Peng ZR, Debette J, Blondeau R (1987) Studies on nutritional factors affecting growth and competition ofPseudomonas maltophilia 1 in the presence ofPseudomonas fluorescens. Syst Appl Microbiol 9:307–311

    Google Scholar 

  29. Stanier RY, Palleroni NJ, Doudoroff M (1966) The aerobic pseudomonads: a taxonomic study. J Gen Microbiol 43:159–271

    PubMed  Google Scholar 

  30. Swings J, De Vos P, Van Den Mooter M, De Ley J (1983) Transfer ofPseudomonas maltophilia Hugh 1981 to the genusXanthomonas asXanthomonas maltophilia (Hugh 1981) comb. nov. Int J Syst Bacteriol 33:409–413

    Google Scholar 

  31. Van Outryve MF, Gosselé F, Kersters K, Swings J (1988) The composition of the rhizosphere of chicory (Cichorium intybus L. var.foliosum Hegi). Can J Microbiol 34:1203–1208

    Google Scholar 

  32. Webb EC (1984) Enzyme nomenclature New York: Academic Press

    Google Scholar 

  33. Weber K, Pringle JR, Osborn M (1972) Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol 26:3–27

    PubMed  Google Scholar 

  34. Whooley MA, O'Callaghan JA, Mc Loughlin AJ (1983) Effect of substrate on the regulation of exoprotease production byPseudomonas aeruginosa ATCC 10145. J Gen Microbiol 129:981–988

    PubMed  Google Scholar 

  35. Wretlind B, Wadström T (1977) Purification and properties of a protease with elastase activity fromPseudomonas aeruginosa. J Gen Microbiol 103:319–327

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Debette, J. Isolation and characterization of an extracellular proteinase produced by a soil strain ofXanthomonas maltophilia . Current Microbiology 22, 85–90 (1991). https://doi.org/10.1007/BF02105381

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02105381

Keywords

Navigation