Statistical modelling of eugenol benzoate synthesis using rhizomucor miehei lipase reinforced nanobioconjugates

Abstract

The chemical route for synthesizing eugenol benzoate is concomitant with a myriad of environmentally unfavorable practices viz. the use toxic chemicals, tedious separation process and emancipation of harmful unwanted by-products. In this perspective, an alternative technique utilizing Rhizomucor miehei lipase (RML) immobilized onto activated chitosan-multiwalled carbon nanotubes (RML/CS/MWCNTs) is proposed. The properties and morphology of the RML/CS/MWCNTs were characterized using field emission scanning electron microscopy (FESEM), Fourier-transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The approach of response surface methodology employing the central composite design (CCD) based on four parameters (incubation time, temperature, substrate molar ratio, and enzyme loading) were used to optimize the experimental conditions for the RML/CS/MWCNTs catalyzed synthesis of eugenol benzoate. The study found that the high yield production of eugenol benzoate is greatly affected by factors such as temperature and incubation time. Under optimized conditions, the highest yield of eugenol benzoate was obtained (56.13%) at 60 °C, 6 h of incubation time, enzyme loading (15 mg) and molar ratio of eugenol/benzoic acid of 4:1. Therefore, the RML/CS/MWCNTs developed here appear to be a promising alternative yet environmentally friendly biocatalyst for a sustainable production of eugenol benzoate.