Optimization of lipase-catalyzed regioselective acylation of pyridoxine (vitamin B6)

Response surface methodology was successfully applied to optimize lipase-catalyzed regioselective esterification of pyridoxine (PN). Effects of various reaction conditions, including reaction temperature, time, enzyme loading, substrate molar ratio, and water activity, were investigated. A central composite design was employed to search for the optimal conversion of PN. A quadratic polynomial regression model was used for analysis of the experimental data at a 95% level (p < 0.05). The analysis confirmed that the water activity was the most significant factor affecting the conversion of PN. It was also suggested that the conversion was strongly affected by independent variables of temperature, time, substrate molar ratio, and water activity as well as interaction terms of temperature and enzyme loading/substrate molar ratio/water activity, time and enzyme loading/substrate molar ratio, substrate molar ratio, and water activity. The coefficient of determination of the model was found to be 0.963. Three sets of optimum reaction conditions were established, and the verified experimental trials were performed for validating the optimum points. A scale-up experiment was also done under the first set of optimal conditions.