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The goal of the current study is to propose and study an innovative method in synthesizing biocatalyst within a microfluidic reactor and later use it in the esterification of β-sitosterol. A droplet microreactor was specially designed and fabricated using soft-lithography technique and was used for formulation of silica nanoparticles. Sol-gel reaction method was adapted in the microreactor to produce nanoparticles which were used as the support for lipase from  Rhizomucor miehei . Another microfluidics reactor was also designed and fabricated for the immobilization of the Lipase on the silica nanoparticles through a physical adsorption technique producing a biocatalyst. The catalytic efficacy of the biocatalyst was next evaluated by esterifying β-sitosterol (the main component of phytosterol) at two different catalyst loadings and comparing it to free lipase under the same experimental parameters. Samples were taken periodically and diluted with n-hexane before injecting them into gas chromatography for analysis. Spherical silica nanoparticles synthesized from the microreactor have good monodispersity with an average size of 480 nm. The silica nanoparticles produced in the microflow system showed large BET surface area, pore volume, and pore size of 125.08 m 2 /g, 0.42 cm 3 /g, and 135.70 Å, respectively.  Rhizomucor miehei  lipase was successfully adsorbed on the support with lipase activity around 16160 U/g. Increasing the catalyst loading increased the esterification rate from 79% to 97.3% when utilized 2.0 and 2.5 mg/ml of immobilized lipase, respectively. The findings also revealed that, under comparable reaction parameters, immobilized lipase had a higher esterification rate than free lipase.

Synthesis of biocatalyst in microfluidic reactor for β-sitosterol esterification