Purification and characterization of a 43.5 kDa keratinolytic metalloprotease from Microsporum canis.

Amino Acid Sequence; Animals; Cat Diseases/microbiology; Cats; Dermatomycoses/microbiology/veterinary; Electrophoresis, Polyacrylamide Gel; Keratins/metabolism; Metalloendopeptidases/antagonists & inhibitors/chemistry/isolation & purification/metabolism; Microsporum/enzymology/growth & development/isolation & purification; Molecular Sequence Data

Abstract :

[en] A keratinolytic protease secreted by a feline clinical isolate of Microsporum canis cultivated in a broth containing feline keratin as the sole nitrogen source was purified from the culture filtrate by affinity chromatography on bacitracin-agarose and by hydrophobic chromatography on octyl-agarose. The enzyme had an apparent molecular mass of 43.5 kDa and the pI was 7.7. It had a significant activity against keratin azure, elastin-Congo red and denatured type I collagen (azocoll). Using the latter substrate, the optimum pH was around 8 and the apparent optimum temperature around 50 degrees C. The protease was strongly inhibited by 1,10-phenanthroline, phosphoramidon and EDTA. The first 13 N-terminal amino acid sequence showed a 61% homology with that of the extracellular metalloprotease of Aspergillus fumigatus and with the neutral protease I of A. oryzae, confirming that this 43.5 kDa keratinase is a metalloprotease. This keratinolytic metalloprotease could be a virulence-related factor involved in pathophysiological mechanisms of M. canis dermatophytosis.

Disciplines :

Veterinary medicine & animal health

Author, co-author :

Brouta, F.

Descamps, F.

Fett, Thomas ; Université de Liège - ULiège > Département de morphologie et pathologie > Pathologie spéciale et autopsies

Losson, Bertrand ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Gerday, C.

Mignon, Bernard ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Language :

English

Title :

Purification and characterization of a 43.5 kDa keratinolytic metalloprotease from Microsporum canis.

Publication date :

2001

Journal title :

Medical Mycology

ISSN :

1369-3786

eISSN :

1460-2709

Publisher :

BIOS Scientific Publishers, Oxford, United Kingdom

Volume :

Issue :

Pages :

269-275

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 12 February 2010

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