Synthesis, characterization and kinetic analysis of chitosan coated magnetic nanobiocatalyst and its application on glucose oleate ester synthesis

Publication date: Available online 16 February 2016 Source:Journal of Molecular Catalysis B: Enzymatic Author(s): Manisha Jain, A.Mariya Sebatini, P. Radha, S. Kiruthika, C. Muthukumaran, K. Tamilarasan In this work, we report the covalent immobilization of lipase enzyme on the surface of chitosan coated magnetic nanoparticles (MNPs) using N-(3-dimethylaminoproyl)-N-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) as coupling agents. Surface functional groups of chitosan coated MNPs, lipase enzyme, and immobilized lipase enzyme were characterized by Fourier Transform Infrared Spectroscopic (FT-IR) analysis. The structural characteristics of the chitosan coated magnetic nanoparticles were analyzed by X-ray Diffraction (XRD) studies. The statistical methodology with central composite design (CCD) was applied to evaluate the effects of immobilization, including the magnetic nanoparticles, lipase enzyme, pH and immobilization time on the enzyme activity. Based on the statistical analysis, the optimum immobilization conditions of magnetic nanoparticles (40mg), lipase enzyme (40mg), pH (9), and immobilization time (6h), the maximum enzyme activity obtained was 16.94U/mL. The optimum pH and temperature for maximum enzyme activity of immobilized lipase are found to be pH 9 and 40°C respectively. The immobilized lipase exhibited excellent catalytic activity over eight successive cycles andretain64% original stability. The Michaelis-Menten enzyme kinetic studies ...
Source: Journal of Molecular Catalysis B: Enzymatic - Category: Biochemistry Source Type: research