Korean J Med Mycol.  1997 Jun;2(1):51-58.

Purification and Characterisation of Extracellular Proteinase from Trichophyton rubrum

• Kim DH
• Lee YS
• Yoo JI
• Choi YH
• Joo HY
• Kim BS
• Kim KS
• Kim JA

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.

Keyword

Trichophyton rubrum; Serine proteinase

MeSH Terms

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
Collagen Type VI
Gelatin
Peptide Hydrolases
Phenylmethylsulfonyl Fluoride
Serine Proteases
Sodium Dodecyl Sulfate
Virulence Factors