To expand the field of nanomaterial and engineering of enzyme in eco-friendly processes, gold mesoflower (Au-MF) nanostructure was applied for preparation of three series of immobilized lipase (Au-MF/SAM 1-3) through biofunctionalization of surface by Ugi multicomponent reaction. The synthesized Au-MF/SAM 1-3/lipase as unique biocatalysts was confirmed by different analytical tools and techniques. Compared to the free lipase, the Au-MF/SAM 1-3/lipase showed more stability at high temperature and pH. Also, these biocatalysts showed high storage stability and reusability after 2 months and eight cycles, respectively. Moreover, the kinetic behavior was investigated and the results showed a minimal impairment of catalytic activity of immobilized lipase. The kinetic constants of the immobilized lipase, Au-MF/SAM 2/lipase, are Km = 0.37 mM, Vmax = 0.22 mM min-1, and kcat = 154 min-1. The immobilized lipase showed smaller activation energy (Ea) than that of free enzyme, indicating that the immobilized enzyme is less sensitive to temperature. In the following, the biodiesel production from palmitic acid was studied in the presence of Au-MF/SAM 2/lipase as an efficient biocatalyst. The influence of different reaction parameters such as temperature, molar ratio of alcohol to palmitic acid, water content, and lipase amount was deeply investigated.
Keywords: biodiesel production; enzyme engineering; gold mesoflower; lipase; postfunctionalization.