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- Title
Purification and biochemical characterization of an extracellular endoglucanase from the necrotrophic oomycete, Pythium myriotylum Dreschler.
- Authors
Geethu, C.; Nair, R. Aswati
- Abstract
An extracellular endoglucanase (EG) that catalyzes the hydrolysis of carboxy-methyl cellulose (CMC) as substrate was purified to homogeneity from the soft-rot causing oomycete P. myriotylum with maximum EG production observed in presence of 1% (w/v) sucrose. The enzyme designated PmEG was observed to be monomeric with a molecular weight of 78 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Optimal activity of PmEG was determined at pH 5.0 and 25 °C with stability observed at pH extending over acidic to alkaline ranges viz., 3.0-10.0 and thermal stability upto 75 °C for 1 h. Optimal PmEG activity was obtained by addition of metal ions viz., Ca2+, Fe3+, Zn2+, Cu2+, Al3+, and also in presence of DTT and β-mercaptoethanol while it was inhibited by Cr2+. Various organic solvents, surfactants, and the oxidant, H2O2 had little/no effect on PmEG activity reflecting its robustness and potential commercial significance. Kinetic constants of PmEG, Km and Vmax were determined as 1.1 mM and 407 µmol min−1 mg−1 protein, respectively. Glucose was observed to cause mixed non-competitive inhibition of PmEG.
- Subjects
CELLULASE; BIOMARKERS; OOMYCETES; PYTHIUM diseases; SODIUM dodecyl sulfate; POLYACRYLAMIDE gel electrophoresis; METAL ions; MERCAPTOETHANOL
- Publication
Journal of Basic Microbiology, 2014, Vol 54, Issue 12, p1322
- ISSN
0233-111X
- Publication type
Article
- DOI
10.1002/jobm.201400323