Abstract

BACKGROUND: Trichophyton rubrum is the most common dermatophyte isolated from human and has ability to invade the tissues such as stratum comeum, nail and hair. The potential role of proteinases as virulence factors of  T. rubrum has been discussed at length.

OBJECTIVE: As a first step towards assessing its virulence role, we report on the purification and characterization of proteinase from T. rubrum isolate culture filtrates.

METHODS: An extracellular serine proteinase has been purified from culture filtrates of Trichophyton rubrum HP-9 by ultrafiltration, gel filtration chromatography, and affinity column chromatography. Azocoll and keratin azure were employed as the substrates of enzyme activities. Peak of proteolytic activity was analyzed by gelatin co-polymerized gel electrophoresis.

RESULTS: The molecular weight of the purified enzyme was approximately exhibited to 14.0 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and molality of 14.0 kDa proteinase activity was 6.0 and 100 mM, respectively. The activity was inhibited by serine proteinase inhibitor, phenylmethylsulfonyl fluoride (PMSF). The proteinase degraded gelatin, collagen type VI, and keratin from human epidermis but not hemoglobin. 

CONCLUSION: The 14,000 Mr extracellular serine proteinase purified from T. rubrum NP-9 culture filtrates has neutral pH optimum 6.0 and activities against gelatin, collagen type VI, and keratin.

Keywords

Serine proteinase Trichophyton rubrum

KJMM 1997 June;2(1):51-58(8). Epub 2016 February 25
Copyright © 1997 by Korean Journal of Medical Mycology

Language

Korean/English

Author

Dong Han Kim; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Yeong Seon Lee; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Jae Il Yoo; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Yeon Hwa Choi; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Hyung Yeul Joo; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Bong Su Kim; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Ki Sang Kim; Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea
Jeong Aee Kim; Department of Dermatology, Seoul National University College of Medicine, Seoul, Korea

Corresponding

Dong Han Kim, Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea.

Publication history

Acknowledgements

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Dong Han Kim

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Yeong Seon Lee

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Jae Il Yoo

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Yeon Hwa Choi

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Hyung Yeul Joo

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Bong Su Kim

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Ki Sang Kim

Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea

Jeong Aee Kim

Department of Dermatology, Seoul National University College of Medicine, Seoul, Korea

Since epub date 2016 February 25