Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol 15 Issue 1, 2013; Page No.(133-142)

STUDIES ON EXTRACELLULAR PROTEINASE ACTIVITY FROM CURVULARIA VERRUCULOSA CAUSING PHAEOHYPHOMYCOSIS

SHESH RAO NAWANGE 1, 2 ,VARSHA AGLAWE 3 , RUCHIKA YADU 1, 3, RUCHI SETHI GUTCH 1,3 ADITI TIWARI , KALPANA WASWANI, SHANKER MOHAN SINGH AND JAISHREE NAIDU

Abstract

We report two cases of phaeohyphomycosis caused by Curvularia verruculosa and subsequent purification and characterization of proteinase produced by it and establishes its significance in pathogenesis. C. verruculosa was isolated repeatedly from subcutaneous lesion and blood of a 7-year-old male child and in another instance from the blood of a 55-year old diabetic male with cellulitis on Sabouraud’s Dextrose Agar (SDA) medium with 0.5mg/mL chloramphenicol. An exocellular proteinase produced by C. verruculosa in a glucose-peptone broth was purified from lyophilized and dialysed culture filtrate of the fungus by Sephadex G-100 gel filtration and SDS- polyacrylamide gel electrophoresis (SDS-PAGE). The purified enzyme was a homogenous protein that could hydrolyse bovine serum albumin and hemoglobin. The enzyme showed neutral pH optimum and maximum activity at 40oC temperature after 60m of incubation. Enzyme was totally inhibited by EDTA and HgCl2. As the strain of C. verruculosa was isolated from peripheral blood of patients with diabetes and cellulitis, it is concluded that this fungus must be producing natural metalloproteinase in-vivo also, that could utilize hemoglobin and serum albumin as protein source for its nutritional requirement in the blood thus surviving and causing infection in such patients.

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