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目的利用响应面法对在异辛烷中脂肪酶催化合成胆固醇癸二酸单烯酯的反应条件进行优化。方法在35℃,摇床转速为220 r·min-1的条件下,选取反应时间(t=4~15 h)、酶的用量(En=2~10 mg)、底物摩尔比即癸二酸二乙烯酯-胆固醇(Sr=5~10)3个因素作为考察对象,以胆固醇转化生成胆固醇癸二酸单烯酯的程度即胆固醇的酯化率(DE)为考察指标,运用design-expert软件的星点设计功能进行实验设计并对实验结果进行分析。结果经实验数据分析表明,t,Sr和En这3个因素均为影响胆固醇酯化的显著因素。经响应面分析可确定最优值为t=11.47 h、En=7.70 mg、Sr=8.93。该条件下的平均酯化率值达90.56%,与响应面拟合方程的酯化率预测值92.80%吻合良好。结论采用星点设计-响应面法优化胆固醇癸二酸单烯酯的酶促催化合成工艺是有效的,可行的。
Objective To optimize the reaction conditions for the synthesis of cholesteryl sebacate monooleate catalyzed by lipase in isooctane using response surface methodology. Methods The reaction conditions (t = 4 ~ 15 h), the amount of enzyme (En = 2 ~ 10 mg), the molar ratio of substrate to decane di Acid-di-divinyl ester-cholesterol (Sr = 5 ~ 10) as the object of study, the cholesterol esterification rate (DE) was calculated by the degree of cholesterol conversion to mono-ester of cholesterol sebacate. Software design of the star point of the experimental design and analysis of experimental results. Results The experimental data show that t, Sr and En are the three factors that affect cholesterol esterification significantly. The response surface analysis showed that the optimal values were t = 11.47 h, En = 7.70 mg, Sr = 8.93. Under these conditions, the average esterification rate of 90.56%, and the response surface fitting equation of the esterification rate of 92.80% predicted good agreement. Conclusion It is feasible and feasible to optimize the enzymatic synthesis of cholesterol sebacic acid mono-ester by the method of star-point design-response surface methodology.