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Purification and Characterization of Fungal L-Asparaginase.

Nalinakshi, G. R. (2011) Purification and Characterization of Fungal L-Asparaginase. [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: The L-asparaginase (E.C.3.5.1.1) enzyme was purified to homogeneity from Botryodiplodia sp. that was grown on wheat bran by solid-state fermentation. Different purification steps like precipitation, ion exchange using Dowex-50WX8 (cation exchanger) and gel filtration on Sepharose 6B column purified the crude filtrate to obtain a pure enzyme preparation. The enzyme was purified to 8.46 folds with a specific activity of 83.3 U. SDS-PAGE of the purified enzyme revealed with a Molecular weight of 160 kDa. The purity was tested by HPLC and capillary electrophoresis. A Lineweaver-Burk and Eadie-Hofstee plot analysis showed a Km value of 0.01M , 0.095M and Vmax of 4 μmol ml-1min-1 and 3.8 μmol ml-1min-1 respectively. The enzyme showed maximum activity at pH 6 when incubated at 30°C for 30 min. L-asparaginase activity was inhibited by PMSF, IA and L-cysteine (Cys).
Uncontrolled Keywords: SSF, L-asparaginase, Botryodiplodia sp., cation exchanger, HPLC
Subjects: 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology > 04 Fungi
600 Technology > 05 Chemical engineering > 03 Enzyme Biotechnology and Engineering
Divisions: Fermentation Technology and Bioengineering
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 01 Aug 2011 11:20
Last Modified: 28 Dec 2011 10:27
URI: http://ir.cftri.com/id/eprint/10344

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