Homolka J
Department of Clinical Biochemistry, Faculty of General Medicine, Charles University, Prague, Czechoslovakia.
J Clin Chem Clin Biochem. 1987 Nov;25(11):805-9. doi: 10.1515/cclm.1987.25.11.805.
The proposed system of continuous monitoring of enzyme activities is based primarily on the electrochemical behaviour of thiol compounds, and the experimental equipment is extremely simple. The determination of cholinesterase (EC 3.1.1.8) activity is described. The normal values obtained for men (73.9, s +/- 10.3 microkat/l) and for women (71.1, s +/- 10.2 microkat/l), lie within the usual range of analogous photometric methods. Systems for determination of the activities of alkaline phosphatase (EC 3.1.3.1) and adenosylhomocysteinase (EC 3.3.1.1) are described. The activity of aspartate aminotransferase (EC 2.6.1.1) is determined by a combination of enzyme reactions, in which CoA is released from acetyl-CoA. Analogous procedures are discussed for determinations of alanine aminotransferase (EC 2.6.1.2), lactate dehydrogenase (EC 1.1.1.27), lipase (EC 3.1.1.2), and phospholipase A2 (EC 3.1.1.4) activities, and for determination of substrates, e.g., acetate and carnitine. Possible determinations of an additional 199 enzyme activities and of some of substrates are suggested. By determining electrochemically active groups other than thiols this method becomes almost universally applicable.
所提出的酶活性连续监测系统主要基于硫醇化合物的电化学行为,并且实验设备极其简单。文中描述了胆碱酯酶(EC 3.1.1.8)活性的测定方法。男性(73.9,s±10.3微卡特/升)和女性(71.1,s±10.2微卡特/升)获得的正常数值,处于类似光度法的通常范围内。文中还描述了碱性磷酸酶(EC 3.1.3.1)和腺苷同型半胱氨酸酶(EC 3.3.1.1)活性的测定系统。天冬氨酸转氨酶(EC 2.6.1.1)的活性通过一系列酶促反应来测定,在这些反应中辅酶A从乙酰辅酶A中释放出来。对于丙氨酸转氨酶(EC 2.6.1.2)、乳酸脱氢酶(EC 1.1.1.27)、脂肪酶(EC 3.1.1.2)和磷脂酶A2(EC 3.1.1.4)活性的测定,以及底物(如乙酸盐和肉碱)的测定,讨论了类似的程序。文中还提出了另外199种酶活性以及一些底物的可能测定方法。通过测定除硫醇以外的电化学活性基团,该方法几乎变得普遍适用。