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. 1989 Nov;57(11):3479–3483. doi: 10.1128/iai.57.11.3479-3483.1989

Isolation of a keratinolytic proteinase from Trichophyton mentagrophytes with enzymatic activity at acidic pH.

R Tsuboi 1, I Ko 1, K Takamori 1, H Ogawa 1
PMCID: PMC259856  PMID: 2478474

Abstract

A keratinolytic proteinase with enzyme activity at acidic pH was isolated from culture filtrates of Trichophyton mentagrophytes, a major pathogenic fungus of dermatophytosis. The molecular weight of the proteinase was estimated to be 41,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 38,000 by gel filtration. The isoelectric point was determined to be 3.9. The proteinase had a pH optimum of 4.5 for keratin and 5.5 for hemoglobin. This enzyme hydrolyzed the synthetic chymotrypsin substrate Suc-Ala-Ala-Pro-Phe-MCA (Km, 0.59 mM), and its activity was strongly inhibited by chymostatin. Previously reported proteinases from dermatophytes have had enzyme activities in neutral or alkaline pH; however, healthy skin has a weakly acidic pH. Thus, the purified proteinase which has an optimal activity at acidic pH and hydrolyzes skin constituents could be an important virulence factor in dermatophytosis.

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Selected References

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  1. Asahi M., Lindquist R., Fukuyama K., Apodaca G., Epstein W. L., McKerrow J. H. Purification and characterization of major extracellular proteinases from Trichophyton rubrum. Biochem J. 1985 Nov 15;232(1):139–144. doi: 10.1042/bj2320139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Day W. C., Toncic P., Stratman S. L., Leeman U., Harmon S. R. Isolation and properties of an extracellular protease of Trichophyton granulosum. Biochim Biophys Acta. 1968 Nov 19;167(3):597–606. doi: 10.1016/0005-2744(68)90050-8. [DOI] [PubMed] [Google Scholar]
  3. Hattori M., Yoshiura K., Negi M., Ogawa H. Keratinolytic proteinase produced by Candida albicans. Sabouraudia. 1984;22(3):175–183. [PubMed] [Google Scholar]
  4. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  5. 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]
  6. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  7. Negi M., Tsuboi R., Matsui T., Ogawa H. Isolation and characterization of proteinase from Candida albicans: substrate specificity. J Invest Dermatol. 1984 Jul;83(1):32–36. doi: 10.1111/1523-1747.ep12261656. [DOI] [PubMed] [Google Scholar]
  8. Philpot C. The differentiation of Trichophyton mentagrophytes from T. rubrum by a simple urease test. Sabouraudia. 1967 Feb;5(3):189–193. [PubMed] [Google Scholar]
  9. SUDMAN M. S., SCHMITT J. A. DIFFERENTIATION OF TRICHOPHYTON RUBRUM AND TRICHOPHYTON MENTAGROPHYTES BY PIGMENT PRODUCTION. Appl Microbiol. 1965 Mar;13:290–290. doi: 10.1128/am.13.2.290-290.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sanyal A. K., Das S. K., Banerjee A. B. Purification and partial characterization of an exocellular proteinase from Trichophyton rubrum. Sabouraudia. 1985 Jun;23(3):165–178. doi: 10.1080/00362178585380271. [DOI] [PubMed] [Google Scholar]
  11. Sawada H., Yokosawa H., Hoshi M., Ishii S. Ascidian sperm chymotrypsin-like enzyme; participation in fertilization. Experientia. 1983 Apr 15;39(4):377–378. doi: 10.1007/BF01963132. [DOI] [PubMed] [Google Scholar]
  12. Takiuchi I., Higuchi D., Sei Y., Koga M. Isolation of an extracellular proteinase (keratinase) from Microsporum canis. Sabouraudia. 1982 Dec;20(4):281–288. [PubMed] [Google Scholar]
  13. Takiuchi I., Higuchi D. [Isolation, purification and biochemical properties of keratinase elaborated from Microsporum gypseum (author's transl)]. Nihon Hifuka Gakkai Zasshi. 1977 Apr;87(5):305–309. [PubMed] [Google Scholar]
  14. Yu R. J., Harmon S. R., Blank F. Hair digestion by a keratinase of Trichophyton mentagrophytes. J Invest Dermatol. 1969 Aug;53(2):166–171. [PubMed] [Google Scholar]
  15. Yu R. J., Harmon S. R., Blank F. Isolation and purification of an extracellular keratinase of Trichophyton mentagrophytes. J Bacteriol. 1968 Oct;96(4):1435–1436. doi: 10.1128/jb.96.4.1435-1436.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yu R. J., Harmon S. R., Grappel S. F., Blank F. Two cell-bound keratinases of Trichophyton mentagrophytes. J Invest Dermatol. 1971 Jan;56(1):27–32. doi: 10.1111/1523-1747.ep12291869. [DOI] [PubMed] [Google Scholar]
  17. Yu R. J., Harmon S. R., Wachter P. E., Blank F. Amino acid composition and specificity of a keratinase of Trichophyton mentagrophytes. Arch Biochem Biophys. 1969 Dec;135(1):363–370. doi: 10.1016/0003-9861(69)90551-7. [DOI] [PubMed] [Google Scholar]
  18. Zimmerman M., Ashe B., Yurewicz E. C., Patel G. Sensitive assays for trypsin, elastase, and chymotrypsin using new fluorogenic substrates. Anal Biochem. 1977 Mar;78(1):47–51. doi: 10.1016/0003-2697(77)90006-9. [DOI] [PubMed] [Google Scholar]

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