固定化β-羟丁酸脱氢酶的研究  

Studies on Immobilization of β-Hydroxybutyrate Dehydrogenase

作　　者：张颖[1] 徐兆珍[1] 于永光[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院体检中心,黑龙江哈尔滨150001

出　　处：《中国实验诊断学》2008年第12期1496-1498,共3页Chinese Journal of Laboratory Diagnosis

基　　金：黑龙江省科学技术厅攻关课题项目(NO.200220101)

摘　　要：目的研究固定化β-羟丁酸脱氢酶的性质及其临床应用价值。方法以壳聚糖为载体,使用戊二醛为交联剂共价键将β-羟丁酸脱氢酶固定化。结果酶蛋白偶联率为60.9%,酶的活性回收率为55.4%;原酶和固定化酶的km值分别为0.8×10-2g/ml和0.5×10-2g/ml;原酶最适温度为37℃,固定化酶最适温度为50℃;原酶最适pH值为8.0左右,固定化酶最适pH值为7.3左右。结论由于固定化酶具有较高的热稳定性(最适50℃)和低温稳定性(4℃8个月),可在较大的温度范围和较长时间内反复使用,有利于环保和降低试剂盒的成本。Objective To study the clinical application of Immobilization of β-Hydroxybutyrate Dehydrogenase on chitosan. Methods Immobilization of β-Hydroxybutyrate Dehydrogenase supporting onto chitosan, cross-linked by glutaraldehyde. Results Usually cross-linked by the rate of coupling enzymatic proteid 60.9 %, the rate of enzyme actively recovery is 55.4%. The km of original enzyme and immobilized enzyme is 0.8 × 10^-2g/ml and 0.5 × 10^-2g/ml.The optimum temperature of original enzyme is 37℃, immobilized enzyme 50℃. The optimum pH of original enzyme is about 8.0, immobilized enzyme is about 7.3. Conclusion Immobilized enzyme has higher hot stability （the most suitable temperature is 50℃ ）and lower temperature stability （ 4℃ ,8 months） ,and can be used repeatedly. It is advanced to environment protecting and to reduce the cost of the reagent kit.

关 键 词：固定化酶 壳聚糖 β-羟丁酸脱氢酶 

分 类 号：R446.1[医药卫生—诊断学]