Facile fabrication of lipase to amine functionalized gold nanoparticles to enhance stability and activity

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

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Official URL: http://dx.doi.org/10.1039/C7RA06075K


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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