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Purification and characterization of an extracellular 24 kDa chitobiosidase from the mycoparasitic fungus Trichoderma saturnisporum

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Title Purification and characterization of an extracellular 24 kDa
chitobiosidase from the mycoparasitic fungus Trichoderma
saturnisporum
 
Creator Sharma, V
Shanmugam, V
 
Subject Plant sciences
Synthetic Chemistry
 
Description A Trichoderma saturnisporum Hamill isolate GITX-Panog (C) exhibiting strong chitinolytic and
antifungal activity against Fusarium oxysporum f.sp. dianthi, the causal agent of vascular wilt in
carnation was used to purify extracellular chitobiosidase using Czapek-Dox broth amended
with the fungal mycelium as the carbon source. The protein was purified by precipitation with
ammonium sulphate, followed by DEAE-Cellulose anion-exchange and Sephacryl S-200 high
resolution gel filtration chromatography. The purity of the enzyme was determined by SDSPAGE,
with an estimated molecular mass of 24 kDa. In native gel assay with 4-methylumbelliferyl-
N,N′ diacetyl-β-D-chitobioside (4-Mu-(GluNAc)2, the purified chitobiosidase was visualized
as single fluorescent band. Enzyme activity towards short oligomeric natural substrates
indicated that the enzyme has properties that are characteristic to exochitinases. The enzyme
was active up to 60 °C and at pH 4.0, and displayed maximum stability at 50 °C. Mn2+ and Zn2+
stimulated the enzyme activity by 63% and 41%, respectively. The Km and Vmax values of the
purified enzyme for 4-Mu-(GluNAc)2 were 338.9 μM ml–1 and 0.119 μM ml–1 min–1, respectively.
This appears to be the first report of characterization of a chitobiosidase from antagonistic
Trichoderma saturnisporum
 
Date 2011
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/1054/1/72_2011_Purification.pdf
Sharma, V and Shanmugam, V (2011) Purification and characterization of an extracellular 24 kDa chitobiosidase from the mycoparasitic fungus Trichoderma saturnisporum. Journal of Basic Microbiology, 51. pp. 1-8.
 
Relation http://ihbt.csircentral.net/1054/