Purification and Characterisation of Extracellular Proteinase from Trichophyton rubrum.
- Author:
Dong Han KIM
1
;
Yeong Seon LEE
;
Jae Il YOO
;
Yeon Hwa CHOI
;
Hyung Yeul JOO
;
Bong Su KIM
;
Ki Sang KIM
;
Jeong Aee KIM
Author Information
1. Laboratory of Nosocomial Pathogens, Department of Microbiological Diseases, National Institute of Health, Korea.
- Publication Type:Original Article
- Keywords:
Trichophyton rubrum;
Serine proteinase
- MeSH:
Arthrodermataceae;
Chromatography;
Chromatography, Gel;
Collagen Type VI;
Electrophoresis;
Electrophoresis, Polyacrylamide Gel;
Epidermis;
Gelatin;
Hair;
Humans;
Hydrogen-Ion Concentration;
Molecular Weight;
Peptide Hydrolases;
Phenylmethylsulfonyl Fluoride;
Serine Proteases;
Sodium Dodecyl Sulfate;
Trichophyton*;
Ultrafiltration;
Virulence;
Virulence Factors
- From:Korean Journal of Medical Mycology
1997;2(1):51-58
- CountryRepublic of Korea
- Language:Korean
-
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 F rMSrMm 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 100mM, 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.
