Interfacing proteins with nanostructured materials offer the possibility to obtain novel bioconjugates for a variety of applications. We report herein the ability of nanostructured poly-D,L-lactic acid (PDLLA) based carriers to enhance protein stability to a greater extent than conventional flat supports with the same chemical structure. PDLLA was processed using an innovative patented methodology that permitted to obtain spherical nanoparticles with a mean diameter of 200 nm. PDLLA nanoparticles were used as carrier for the physical adsorption of Candida rugosa lipase preparations (CRL). Enzymatic activity and stability before and after conjugation to the nanopolymeric support was evaluated in different conditions (pH, T, organic solvents). Also, FTIR characterization was employed for free and immobilized enzymes. Immobilization on nanocarriers affected significantly CRL performance, leading to higher specific activity and stability to denaturing agents. FTIR measurements showed a reproducible modification in the conformational features for the CRL in different media (solid state and aqueous solutions). In an attempt to assess the structure-performance relationship of the immobilized CRL based on the kinetic studies together with the FTIR characterization, we propose that the enhancement of the enzyme performance is closely related to the conformational changes of the protein structure due to the immobilization on nanocarriers.

Structure-Performance Relationships of Lipolytic Enzymes Immobilized on Polymeric Nanoparticles / Chronopoulou, Laura; G., Kamel; Bordi, Federico; M., Diociaiuti; Lupi, Stefano; Palocci, Cleofe. - STAMPA. - (2010). (Intervento presentato al convegno From Solid State to Biophysics: from Physics to Life Sciences tenutosi a Cavtat, Dubrovnik, Croatia nel 12-19/06/2010).

Structure-Performance Relationships of Lipolytic Enzymes Immobilized on Polymeric Nanoparticles

CHRONOPOULOU, LAURA;BORDI, FEDERICO;LUPI, Stefano;PALOCCI, Cleofe
2010

Abstract

Interfacing proteins with nanostructured materials offer the possibility to obtain novel bioconjugates for a variety of applications. We report herein the ability of nanostructured poly-D,L-lactic acid (PDLLA) based carriers to enhance protein stability to a greater extent than conventional flat supports with the same chemical structure. PDLLA was processed using an innovative patented methodology that permitted to obtain spherical nanoparticles with a mean diameter of 200 nm. PDLLA nanoparticles were used as carrier for the physical adsorption of Candida rugosa lipase preparations (CRL). Enzymatic activity and stability before and after conjugation to the nanopolymeric support was evaluated in different conditions (pH, T, organic solvents). Also, FTIR characterization was employed for free and immobilized enzymes. Immobilization on nanocarriers affected significantly CRL performance, leading to higher specific activity and stability to denaturing agents. FTIR measurements showed a reproducible modification in the conformational features for the CRL in different media (solid state and aqueous solutions). In an attempt to assess the structure-performance relationship of the immobilized CRL based on the kinetic studies together with the FTIR characterization, we propose that the enhancement of the enzyme performance is closely related to the conformational changes of the protein structure due to the immobilization on nanocarriers.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/384951
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