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

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Facile fabrication of lipase to amine functionalized gold nanoparticles to enhance stability and activity
 
Creator Sristy, Shikha
Thakur, Krishan Gopal
Bhattacharyya, Mani Shankar
 
Subject QR Microbiology
 
Description 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.
 
Publisher RSC
 
Date 2017
 
Type Article
PeerReviewed
 
Relation http://dx.doi.org/10.1039/C7RA06075K
http://crdd.osdd.net/open/2080/
 
Identifier 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