Enhanced Activity and Stability of Cellobiase (b-Glucosidase: EC 3.2.1.21) Produced in the Presence of 2-deoxy-D-glucose from the Fungus Termitomyces Clypeatus
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
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| Title |
Enhanced Activity and Stability of Cellobiase (b-Glucosidase: EC 3.2.1.21) Produced in the Presence of 2-deoxy-D-glucose from the Fungus Termitomyces Clypeatus
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| Creator |
Ghorai, Shakuntala
Chowdhury, Sudeshna Pal, Swagata Banik, Samudra Prasad Mukherjee, Sumana Khowala, Suman |
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| Subject |
Drug Development/Diagnostics & Biotechnology
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| Description |
Generally less glycosylation or deglycosylation has a detrimental effect on enzyme activity and stability.
Increased production and secretion of cellobiase was earlier obtained in the presence of the glycosylation
inhibitor 2-deoxy-D-glucose in filamentous fungus Termitomyces clypeatus [Mukherjee, S.; Chowdhury, S.;
Ghorai, S.; Pal, S.; Khowala, S. Biotechnol. Lett. 2006, 28, 1773–1778]. In this study the enzyme was purified
from the culture medium by ultrafiltration and gel-permeation, ion-exchange and high-performance
liquid chromatography, and its catalytic activity was six times higher compared to the control enzyme.
Km and Vmax of the purified enzyme were measured as 0.187 mM and 0.018 U mg�1, respectively, using
pNPG as the substrate. The enzyme had temperature and pH optima at 45 �C and pH 5.4, respectively,
and retained full activity in a pH range of 5–8 and temperatures of 30–60 �C. Interestingly less glycosylated
cellobiase was resistant towards proteolytic as well as endoglycosidase-H digestion and showed
higher stability than native enzyme due to increased aggregation of the protein. The enzyme also showed
higher specific activity in the presence of cellobiose and pNPG and less susceptibility towards salts and
different chemical agents. The b-glucosidase can be considered as a potentially useful enzyme in various
food-processing, pharmaceutical and fermentation industries.
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| Date |
2010
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/993/1/47_carbohydrate_2010.pdf
Ghorai, Shakuntala and Chowdhury, Sudeshna and Pal, Swagata and Banik, Samudra Prasad and Mukherjee, Sumana and Khowala, Suman (2010) Enhanced Activity and Stability of Cellobiase (b-Glucosidase: EC 3.2.1.21) Produced in the Presence of 2-deoxy-D-glucose from the Fungus Termitomyces Clypeatus. Carbohydrate Research, 345 (8). pp. 1015-1022. |
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| Relation |
http://dx.doi.org/10.1016/j.carres.2010.02.021
http://www.eprints.iicb.res.in/993/ |
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