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Use of Affinity Matrix for the purification of enzymes
Srividya Chandramouli, R. (2004) Use of Affinity Matrix for the purification of enzymes. [Student Project Report]
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Abstract
This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.
| Item Type: | Student Project Report |
|---|---|
| Additional Information: | Affinity precipitation of enzymes offers a high degree of specificity and provides an efficient extraction of a single product from a crude mixture of proteins, typically found in bioprocessing. Sodium alginate is useful in affinity precipitation as it can be used as a reversible soluble-insoluble macroligand for the affinity precipitation of amylase from Aspergillus oryzae. It was possible to elute amylase from alginate beads. The enzyme was homogenous with a molecular weight of 47 KDa . The native enzyme (crude preparation) has pH optimum of 5.5 and a temperature optimum of 35ºC. The enzyme lost 50% of its activity when heated to 55ºC, for 15 min. the entrapped amylase in alginate had a more alkaline pH optimum of 6.3. It was active over the temperature range of 40-55ºC, with an optimum of 50ºC. Thus, there was a shift of 15ºC, towards higher temperature. Enzyme stability was also higher with 50% of activity retained even after heating at 65ºC, for 15 minutes. Hence, affinity precipitation is an attractive alternative for both recovery of pure enzyme as well as for immobilization of the enzyme. |
| Uncontrolled Keywords: | alpha-amylase, affinity precipitation, alginate beads, purification of enzymes. |
| Subjects: | 500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 16 Enzyme Chemistry |
| Divisions: | Human Resource Development |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 08 Jun 2005 |
| Last Modified: | 28 Dec 2011 09:24 |
| URI: | http://ir.cftri.res.in/id/eprint/44 |
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