Abstract
A new approach was used for the immobilization of Thermomyces lanuginosus lipase (TLL), Candida antarctica lipase B (CALB), and Rhizomucor miehei lipase (RML) on amine-functionalized magnetic nanoparticles (Fe3O4@SiO2-NH2) via a multi-component reaction route (using cyclohexyl isocyanide). The used method offered a single-step and very fast process for covalent attachment of the lipases under extremely mild reaction conditions (25 °C, water, and pH 7.0). Rapid and simple immobilization of 20 mg of RML, TLL, and CALB on 1 g of the support produced 100%, 98.5%, and 99.2% immobilization yields, respectively, after 2 h of incubation. The immobilized derivatives were then used for biodiesel production from waste cooking oil. Response surface methodology (RSM) in combination with central composite rotatable design (CCRD) was employed to evaluate and optimize the biodiesel production. The effect of some parameters such as catalyst amount, reaction temperature, methanol concentration, water content for TLL or water-adsorbent for RML and CALB, and ratio of t-butanol (wt%) were investigated on the fatty acid methyl esters (FAME) yield.