辣根过氧化物酶酶标抗原催化α-萘酚微量热

Microcalorimetric Study of Enzymatic Reaction of α-Naphthol Using Horseradish Peroxidase Labeled Clenbuterol Antigen

采用戊二醛法将盐酸克伦特罗(CL)和辣根过氧化物酶(HRP)偶联制备不同偶联率的酶标抗原,利用MicroDSCⅢ微量热仪测定不同偶联率的酶标抗原催化α-萘酚放热量。结果表明:采用戊二醛法进行偶联HRP的酶活损失微弱,且在相同CL浓度下偶联率低的酶标抗原放热量较大。在同等条件下,0.15mol/LNaCl反应体系中酶促反应的放热量优于0.1mol/LPBS反应体系,在0.15mol/LNaCl缓冲体系下,酶活力为2U时催化放热的最佳底物浓度为20mmol/L。

Clenbuterol （CL） was conjugated to horseradish peroxidase using glutaraldehyde at different ratios and a Micro DSC Ⅲ calorimeter was adopted to measure energy released during hydrolysis ofα-naphthol by horseradish peroxidase labeled CL antigen with different conjugation ratios. A slight decrease was found in enzyme activity and the enzyme labeled antigen with lower conjugation degree showed a higher energy change under the same CL concentration. Under the same condition, the energy released during enzymatic reaction in 0.15 mol/L NaC1 buffer was higher than that in 0.1 mol/L PBS buffer. In 0.15 mol/L NaCI buffer, the optimal concentration of a-naphthol was 20 mmol/L when the enzyme activity was 2 U.