IMMOBILIZATION OF FIREFLY LUCIFERASE ON GLASS RODS - PROPERTIES OF IMMOBILIZED ENZYME
IMMOBILIZATION OF FIREFLY LUCIFERASE ON GLASS RODS - PROPERTIES OF IMMOBILIZED ENZYME
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DOI：
10.1016/0003-2697(77)90672-8
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发表时间：
1977-01-01
影响因子：
2.9
通讯作者：
DELUCA, M
中科院分区：
文献类型：
--
作者：
LEE, Y;JABLONSKI, I;DELUCA, M
用戊二醛将萤火虫荧光素酶共价连接到已粘合到玻璃棒上的烷基胺玻璃珠上。固定化酶具有比可溶性酶低的最适pH，并且发射在615 nm处具有主峰的光，而可溶性酶发射在562 nm处具有峰的光。固定化酶稳定性好，可用于多种检测。峰值光强度在1 × 10 - 4范围内与ATP浓度成线性关系。10-5到1 ×10-8 M.荧光素酶棒用于偶联测定，以测量肌酸磷酸激酶催化的ATP产生速率。这种固定化的荧光素酶对于测定任何类型样品中的低水平ATP都是非常有用的。
Firefly luciferase was covalently linked with glutaraldehyde to alkylamine glass beads which had been cemented to glass rods. The immobilized enzyme has a lower pH optima than the soluble enzyme and emits light with a major peak at 615 nm, while the soluble enzyme emits light with a peak at 562 nm. The immobilized enzyme is stable and can be used for multiple assays. The peak light intensity is linear with respect to ATP concentration in the range of 1 .times. 10-5 to 1 .times. 10-8 M. The luciferase rods were used in a coupled assay to measure the rate of ATP production catalyzed by creatine phosphokinase. This immobilized luciferase should be very useful for assaying low levels of ATP in any type of sample.