Construction of multi-chimeric pyrroloquinoline quinone glucose dehydrogenase with improved enzymatic properties and application in glucose monitoring
Construction of multi-chimeric pyrroloquinoline quinone glucose dehydrogenase with improved enzymatic properties and application in glucose monitoring
复制标题
具有改进酶学性质的多嵌合吡咯喹啉醌葡萄糖脱氢酶的构建及其在血糖监测中的应用
DOI：
10.1023/a:1005658316948
复制
发表时间：
2000
影响因子：
2.7
通讯作者：
K. Sode
中科院分区：
文献类型：
--
作者：
H. Yoshida;T. Iguchi;K. Sode
将大肠杆菌和醋酸钙不动杆菌吡咯喹啉葡萄糖脱氢酶(PQQGDH)决定酶性质的蛋白质区域结合，构建了多嵌合酶。与野生型相比，构建的多嵌合PQQGDH表现出更高的辅因子结合稳定性、热稳定性、底物专一性的改变和葡萄糖的Km值10倍的增加。观察了每个引入的蛋白质区域对酶性质改善的累积效应。将多嵌合PQQGDH应用于电流型葡萄糖传感器的构建，实现了更大的动态范围、更高的操作稳定性和更窄的底物专属性。该葡萄糖传感器可以测量5到40微米的血糖，这表明它有可能直接测量糖尿病患者的高血糖水平。
A multi-chimeric enzyme was constructed by combining the protein regions responsible for the enzymatic properties ofEscherichia coliandAcinetobacter calcoaceticuspyrroloquinoline quinone glucose dehydrogenase (PQQGDH). The constructed multi-chimeric PQQGDH showed increased co-factor binding stability, thermal stability, an alteration in substrate specificity and a 10-fold increase in theKmvalue for glucose compared with the wild-typeE. coliPQQGDH. The cumulative effect of each introduced protein region on the improvement of enzymatic properties was observed. The application of the multi-chimeric PQQGDH in amperometric glucose sensor construction achieved an expanded dynamic range together with increased operational stability and narrower substrate specificity. The glucose sensor can measure glucose from 5 to 40 mM, suggesting its potential for the direct measurement of high blood-glucose levels in diabetic patients.