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本文采用响应面分析法对树脂NKA-Ⅱ固定脂肪酶的条件进行优化,并对其酶学性质和在催化光皮树油合成生物柴油中的操作稳定性进行了简单的探讨。首先通过单因素试验研究酶液/载体比、温度、缓冲液pH值等条件对固定化脂肪酶酶活力的影响,在此基础上,利用响应面试验设计对树脂NKA-Ⅱ固定化脂肪酶的条件进行优化,用制备的固定脂肪酶催化光皮树油和甲醇反应合成生物柴油。结果表明:酶液/载体比、温度和缓冲液pH值对固定化酶的活力都有影响,树脂NKA-Ⅱ固定化脂肪酶的最佳固定条件是:酶液/载体比为4∶1、温度为39℃、缓冲液pH值为7.5,固定化脂肪酶湿重活力高达216.2±4.5U/g,生物柴油转化率达90%以上。将响应面法应用于树脂NKA-Ⅱ固定化脂肪酶条件的优化是可行的,制备的固定脂肪酶具有工业化生产的潜力。
In this paper, the conditions of resin immobilized NKA-Ⅱ lipase were optimized by response surface methodology, and its enzymatic properties and operational stability in catalysis of Biodiesel from Catharanthus roseus were briefly discussed. Firstly, the effect of enzyme liquid / carrier ratio, temperature and buffer pH value on the activity of immobilized lipase was studied by single factor experiment. On the basis of this, the response surface of NKA-Ⅱ immobilized lipase Conditions were optimized, and the prepared immobilized lipase was used to catalyze the reaction of beech tree oil and methanol to synthesize biodiesel. The results showed that the optimum conditions for the immobilization of resin NKA-Ⅱ immobilized lipase were: enzyme solution / carrier ratio, temperature and buffer pH value on the activity of immobilized enzyme: The temperature was 39 ℃, the pH value of buffer was 7.5, the wet weight of immobilized lipase was 216.2 ± 4.5U / g, and the conversion rate of biodiesel was over 90%. It is feasible to apply the response surface method to the immobilization of resin NKA-Ⅱ in lipase. The prepared immobilized lipase has the potential of industrial production.