Sristy, Shikha and Thakur, Krishan Gopal and Bhattacharyya, Mani Shankar (2017) Facile fabrication of lipase to amine functionalized gold nanoparticles to enhance stability and activity. RSC Advances, 7 (68). pp. 42845-42855. ISSN 2046-2069
Full text not available from this repository. (Request a copy)Abstract
Among various techniques of immobilization, EDC/NHS cross linking is a simple and single step process for covalent coupling between enzymes and nanoparticles. Here we describe immobilization of lipase on amine functionalized gold nanoparticles (AuNPs-NH2) to attain enhanced activity and stability. To achieve a suitable orientation, it is necessary to understand the contribution of different functional groups on the enzyme's surface. Therefore, the crystal structure of lipase was analyzed using a computational method (PyMOL) to find the exposed acidic amino acid residues that can be exploited for conjugation. Confirmation of conjugation (AuNP-NH2-lipase) was determined by various techniques such as agarose gel electrophoresis, zeta measurement, FTIR-spectroscopy and TEM. Further, catalytic parameters (Vmax, KM,app, Kcat, and Kcat/KM,app) have been studied to establish activity enhancement upon immobilization. The data also suggested that, AuNP-NH2-lipase has desirable improved parameters such as temperature and storage stability. The thermodynamic parameters for the kinetics of deactivation (Image ID:c7ra06075k-t1.gif, Image ID:c7ra06075k-t2.gif and Image ID:c7ra06075k-t3.gif) of the AuNP-NH2-lipase and free lipase demonstrated better stability of the conjugate. CD and fluorescence spectroscopic studies revealed minor structural rearrangements in the enzyme upon conjugation. Thus the AuNP-NH2-lipase conjugate represents a novel enzyme preparation with attributes of high activity and stability that could be an attractive choice in diverse applications ranging from catalysis to diagnostics.
| Item Type: | Article |
|---|---|
| Additional Information: | Copyright of this article belongs to RSC. |
| Subjects: | Q Science > QR Microbiology |
| Depositing User: | Dr. K.P.S.Sengar |
| Date Deposited: | 02 Apr 2018 09:22 |
| Last Modified: | 02 Apr 2018 09:38 |
| URI: | http://crdd.osdd.net/open/id/eprint/2080 |
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