Otero C, Ballesteros A, Guisán J M
Instituto de Catálisis y Petroleoquimica, CSIC, Madrid, Spain.
Appl Biochem Biotechnol. 1988 Nov;19(2):163-75. doi: 10.1007/BF02921481.
With the aim of fixing the enzyme to the matrix by multiple covalent linkages, lipase from Candida rugosa (formerly cylindracea) has been insolubilized through its amino groups on Sepharose 6B previously activated with 2,3-epoxy-1-propanol. Two main variables that are known to control the number of bonds formed have been tested: the contact time between enzyme and activated support, and the temperature at which the immobilization reaction is carried out. Studies on activity and stability of the different derivatives prepared showed that higher temperatures and longer contact times lead to insolubilized enzymes that are more resistant to inactivation by temperature and the presence of organic solvents. At 50 degrees C and pH 7.2, the insoluble lipase was found to be 140 times more stable than its soluble counterpart.
为了通过多个共价键将酶固定在基质上，来自皱褶假丝酵母（原柱形假丝酵母）的脂肪酶已通过其氨基固定在先前用2,3-环氧-1-丙醇活化的琼脂糖6B上。已知控制形成键数量的两个主要变量已被测试：酶与活化载体之间的接触时间，以及进行固定化反应的温度。对所制备的不同衍生物的活性和稳定性研究表明，较高的温度和较长的接触时间会导致固定化酶对温度和有机溶剂的失活更具抗性。在50℃和pH 7.2条件下，发现不溶性脂肪酶比其可溶性对应物稳定140倍。
