Botrè C, Botrè F
Dipartimento di Studi di Chimica e Tecnologia, Università La Sapienza, Rome, Italy.
Anal Biochem. 1990 Mar;185(2):254-64. doi: 10.1016/0003-2697(90)90289-l.
A potentiometric method capable of determining carbonic anhydrase (CA) activity in vitro and based on the use of a pCO2 sensor is presented. By means of the procedure described here it is possible to follow the rate of CO2 diffusion that takes place in a buffered solution of NaHCO3 in either the presence or the absence of CA. All experimental parameters that affect the speed of HCO3- dehydration, as well as the speed of CO2 diffusion, can be fixed and kept constant for the duration of every assay. The advantage of this method is that the overall dehydration plus diffusion process can be followed as it actually takes place in open thermodynamic systems far from equilibrium. The results obtained strongly confirm the hypothesis of a facilitating role of CA toward the rate of CO2 diffusion.
本文介绍了一种电位测定法,该方法基于使用pCO2传感器能够在体外测定碳酸酐酶(CA)的活性。通过此处所述的程序,可以跟踪在有或没有CA的情况下,在NaHCO3缓冲溶液中发生的CO2扩散速率。在每次测定过程中,所有影响HCO3-脱水速度以及CO2扩散速度的实验参数都可以固定并保持恒定。该方法的优点是,可以跟踪整个脱水加扩散过程,因为它实际上发生在远离平衡的开放热力学系统中。所获得的结果有力地证实了CA对CO2扩散速率具有促进作用的假设。