Improved Production and Properties of β-Glucosidase Influenced by 2-Deoxy-D-Glucose in the Culture Medium of Termitomyces clypeatus
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Improved Production and Properties of β-Glucosidase Influenced by
2-Deoxy-D-Glucose in the Culture Medium of Termitomyces clypeatus
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| Creator |
Ghorai, Shakuntala
Mukherjee, Sumana Mukherjee, Soumya Khowala, Suman |
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| Subject |
Drug Development/Diagnostics & Biotechnology
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| Description |
Increased production, secretion, and activity of
β-glucosidase in the filamentous fungus Termitomyces
clypeatus was achieved in presence of the glycosylation
inhibitor 2-deoxy-D-glucose (0.05%, w/v) during submerged
fermentation. Enzyme activity increased to 163 U/mL by
adding mannose (2 mg/mL) to the medium. Such a high
enzyme activity has not been achieved without mutation or
genetic manipulation. The Km and Vmax of the enzyme in
culture medium were determined to be 0.092 mM and
35.54 U/mg, respectively, with p-nitrophenyl β-D-glucopyranoside
as substrate, confirming its high catalytic activity.
The enzyme displayed optimum activity at pH 5.4 and
45°C. The enzyme was fairly stable between acidic to
alkaline pH and retained about 75 ~ 65% residual activities
between pH 4 and 10.6 and demonstrated full activity at
45ºC for 3 days. The enzyme was also stable in the
presence of Zn2+ and Mg2+ and 80% of the residual activity
was observed in the presence of Mn2+, Ca2+, K+, Cu2+,
EDTA, and sodium azide. Around 70% of the activity was
retained in the presence of 2 M guanidium HCl and 3 M
urea, whereas the activity was 5 and 2 times higher in the
presence of 4 mM beta-mercaptoethanol and 50 mM DTT,
respectively. The enzyme obtained from the culture filtrate
showed potential cellulose saccharifying ability which
increased further when supplemented with commercial
cellulase. Thus, this enzyme could be used without any
additional downstream processing for commercial cellulase preparation and production of bioethanol or for other biotechnological
applications.
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| Date |
2011
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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/94/1/BIOTECHNOLOGY_AND_BIOPROCESS_ENGINEERING%2C(16)_2%2C297%2D304%2C2011[82].pdf
Ghorai, Shakuntala and Mukherjee, Sumana and Mukherjee, Soumya and Khowala, Suman (2011) Improved Production and Properties of β-Glucosidase Influenced by 2-Deoxy-D-Glucose in the Culture Medium of Termitomyces clypeatus. Biotechnology and Bioprocess Engineering, 16. pp. 297-304. |
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| Relation |
http://dx.doi.org/10.1007/s12257-010-0236-3
http://www.eprints.iicb.res.in/94/ |
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