Abstract
Fe3O4 (magnetite) nanoparticles were prepared by coprecipitation method, coated by chitosan and functionalized by glutaraldehyde. Lipase enzyme from Candida rugosa was immobilized on the prepared particles via cross linking reaction. Synthesis steps and characterization were examined by XRD, TEM, and FTIR. 
 The immobilization conditions were 10 mL of phosphate buffer (0.1 M, pH 6.5) containing 30 mg of functionalized magnetic chitosan nanoparticles and 2.0 mg·mL-1 of lipase, immobilization temperature of 4 ℃ and immobilization time of 1 h. Under these conditions, lipase was successfully immobilized with loading capacity of 87 mg/g.
 The immobilized enzyme showed good operational and storage stability, where it remained stable after 30 days of storage at 4◦C.and retained about 61% of its initial activity after twenty repeated uses. Finally enzymatic catalyze synthesis of butyl and hexyl oleate at 40 ◦C with shaking (200 rpm) was realized in n-hexane and confirmed by GC analysis.
Highlights
INTRODUCTIONLipase (triacylglycerol acylhydrolase, EC 3.1.1.3) is regarded as one of the most important and widely used enzyme in industrial and analytical chemistry applications [1,2,3]
Lipase is regarded as one of the most important and widely used enzyme in industrial and analytical chemistry applications [1,2,3]
Magnetic support was firstly applied in enzyme immobilization in 1973 which allow the magnetic separation of the immobilized enzyme from the reaction system [10]
Summary
Lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) is regarded as one of the most important and widely used enzyme in industrial and analytical chemistry applications [1,2,3]. Enzyme immobilization is an efficient technique to enhance the enzyme stability and to allow its reuse. Fe3O4 (magnetite) nanoparticles are widely used in enzyme immobilization due to its biocompatibility, high surface area to volume ratio [11]. Magnetite nanoparticles (MNP) are commonly superficially modified by coating with organic, inorganic or polymers. Polymeric coatings are characterized by better dispersion and stability in solvents than inorganic or small organic compounds [12]. Chitosan as immobilization support is characterized by widely commercial availability, non-toxicity, and low cost [13]. In the present study lipase from Candida rugosa was immobilized on chitosan coated magnetite nanoparticles. Activity, reusability and stability in both aqueous and organic solvents were investigated
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