Duan_2012_J.Biotechnol_159_44

t-Butanol was an excellent reaction medium for enzyme-mediated esterification of oleic acid with glycerol for 1,3-diolein preparation which has been proved by our group. Herein, to achieve the rational synthesis of 1,3-diolien, response surface methodology was applied to examine the effects of the significant variables and their reciprocal effects on the product synthesis. Under the optimal conditions (62.4 degrees C, 0.75g Novozym 435, substrate molar ratio (oleic acid/glycerol) 2.4 and 4.8g t-butanol), the diolein yield of 87.4% could be achieved, and the value of 1,3-diolien/diolein was as high as 87.8%, which were quite close to the predicted values. Compared with the results of our previous single factor experiment, although the values of diolein yield and 1,3-diolien/diolein could not be improved markedly, the enzyme dosage and the reaction medium were spared by 25% and 20%, respectively, which was a remarkable improvement of the enzymatic process.