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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 10, No. 2, 2007, pp. 260-270
Bioline Code: ej07026
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 10, No. 2, 2007, pp. 260-270

 en Purification and characterization of β-glucosidase from Melanocarpus sp. MTCC 3922
Kaur, Jatinder; Chadha, Bhupinder S.; Kumar, Badhan A.; Ghatora, S. Kaur & Saini, Harvinder S.

Abstract

This study reports the purification and characterization of β-glucosidase from a newly isolated thermophilic fungus, Melanocarpus sp. check for this species in other resources Microbial Type Culture Collection (MTCC) 3922. The molecular weight of β-glucosidase was determined to be ~ 92 and 102 kDa with SDS PAGE and gel filtration, respectively, and pI of ~ 4.1. It was optimally active at 60°C and pH 6.0, though was stable at 50°C and pH 5.0 - 6.0. The presence of DTT, mercaptoethanol and metal ions such as Na+, K+, Ca2+, Mg2+ and Zn2+ positively influenced the activity of β-glucosidase but the activity was inhibited in the presence of CuSO4. β-Glucosidase recognized pNP- β-glucopyranoside (pNPG) as the preferred substrate, and showed very low affinity for pNP- β-D-cellobioside. Km and Vmax for the hydrolysis of pNPG by β-glucosidase was calculated as 3.3 mM and 43.68 μmolmin-1mg protein-1, respectively and kcat was quantified as 4 x 103 min-1. β-Glucosidase activity was enhanced appreciably in the presence of alcohols (methanol and ethanol) moreover, purified β-glucosidase showed putative transglycosylation activity that was positively catalyzed in presence of methanol as an acceptor molecule.

Keywords
b-glucosidase, Melanocarpus sp., purification, substrate specificity, transglycosylation

 
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