脂酶非均相催化反应生物传感器的研究

Study on Biosensor of Heterogeneous Reaction Conjugated by Lipase

研究固定化脂肪酶在生物传感器上的响应,研制了能将脂肪酶催化底物所产生的电子转移过程转换为电流信号输出的电化学生物传感器,酯酶非均相反应体系温度为65℃,pH为7.7时验证脂酶活性的变化规律。测定反应体系电流强度的变化趋势为首先是平稳,反应开始电流逐渐增大,随着甘二酯逐渐增多,电流变化缓慢;当甘一酯的量逐渐增多电流也变大,反应结束电流趋于平稳。用液相色谱仪跟踪测定反应过程中底物的含量,结果显示测量相对误差仅为4.9%。证明利用脂酶非均相催化反应生物传感器测定酶催化底物反应的程度是可行的,为实现计算机连续在线检测提供了理论基础。

In order to study immobilized lipase responded in biosensor,it had developed an electrochemical biosensor which could transform the electron metastasis processes produced by lipase-conjugated substrate to current signal output,and the electron metastasis of the lipase in the lipase heterogeneous reaction system was verified at 65 ℃,pH 7.7.The trends in current intensity of reaction was determined as follows： it was stable firstly,then gradually increased with the reaction starting,and then changed slowly along with the increase of monoglyceride;when monoglyceride gradually increased,current also became greater,and the current became stable when reaction was to the end.Liquid chromatography was used to determine the content of the substrate of the reaction.The results showed that the relative error was only 4.9%.It proved that it was feasible to determine the extent of the reaction conjugated by enzyme with the biosensor of lipase heterogeneous conjugating reaction,so as to provide a basic premise for realizing the continuous detection on-line by a computer.