IMMOBILIZATION OF CANDIDA-RUGOSA LIPASE AND SOME PROPERTIES OF THE IMMOBILIZED ENZYME
IMMOBILIZATION OF CANDIDA-RUGOSA LIPASE AND SOME PROPERTIES OF THE IMMOBILIZED ENZYME
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DOI：
10.1016/0141-0229(93)90144-q
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发表时间：
1993-03-01
影响因子：
3.4
通讯作者：
SERRA, JL
中科院分区：
文献类型：
--
作者：
MONTERO, S;BLANCO, A;SERRA, JL
将来自Candida rugosa的脂肪酶（三酰甘油酯水解酶，E.C.3.1.1.3）固定在市售微孔聚丙烯上。酶被迅速吸附在支持物上，在室温下孵育1分钟后，超过60%的可溶性活性从培养基中消失。当在固定化方案结束时立即用橄榄油测定湿制剂时，获得了21%的固定化活性的回收率。固定化酶的活性随水的流失而急剧下降。用有机溶剂预处理的支持显着增加回收的固定化活性。我们的研究结果强烈表明，可溶性脂肪酶可以存在于不同的聚集形式取决于介质的pH值。在酸性pH下，高分子量形式的酶的相对比例高于pH 7.0，这表明脂肪酶也将根据固定化过程中使用的pH以不同的聚集形式固定化。交联的吸附酶与戊二醛降低其活性，但增加了对Triton X-100的洗出效果的脂肪酶的稳定性。最相关的催化性能的可溶性和固定化酶，如最佳pH值和温度，以及范围内的稳定性，动力学参数，和活化能的水解橄榄油和对硝基苯基乙酸酯的数据，报告。
Lipase (triacylglycerol ester hydrolase, E.C.3.1.1.3) from Candida rugosa has been immobilized on commercially available microporous polypropylene. The enzyme was rapidly adsorbed on the support, and more than 60% of the soluble activity disappeared from the medium after 1 min of incubation at room temperature. A recovery of immobilized activity of 21% was obtained when the wet preparation was immediately assayed with olive oil at the end of the immobilization protocol. The activity of the immobilized enzyme drastically decreased with the loss of water of the preparation. Pretreatment of the support with organic solvents significantly increased the recovered immobilized activity. Our results strongly suggest that the soluble lipase could exist in different aggregation forms depending on the pH of the medium. At acidic pH, the relative proportion of high-molecular-weight forms of the enzyme is higher than at pH 7.0, suggesting that the lipase would be also immobilized in different aggregation forms depending on the pH used in the immobilization procedure. Crosslinking of the adsorbed enzyme with glutaraldehyde diminished its activity but increased the stability of the lipase against the washing-out effect of Triton X-100. Data on the most relevant catalytic properties of the soluble and immobilized enzyme, such as optimum pH and temperature as well as ranges of stability, kinetic parameters, and activation energy for the hydrolysis of olive oil and p-nitrophenyl acetate, are reported.