Department of Physical Chemistry, Faculty of Chemical Technology, University of Pardubice, Czech Republic.
Bioresour Technol. 2010 Aug;101(15):6281-3. doi: 10.1016/j.biortech.2010.03.017. Epub 2010 Mar 23.
Homological series of 14 achiral derivates and series of five chiral derivates of imidazole were tested in vitro as inhibitors of hydrolysis of acetylcholine using enzyme preparation of acetylcholinesterase from electric eel. The batch stirred reactor at 25 degrees C, pH 8 (phosphate buffer), ionic strength 0.11 M and catalytic activity of the enzyme preparation 0.14 U ml(-1) of the reaction mixture were used. The temporal dependences of actual concentrations of acetylcholine, choline and acetic acid were determined by an original HPLC method. For all used inhibitors, these time dependences conform with the probability of more than 90% to the model of competitive irreversible inhibition. All kinetic constants including k(3) defining the rate of inhibition (0.38-5.3M(-1)s(-1)) and qualified estimation of the absolute acetylcholinesterase concentration in the reaction mixture (40-110 nM) were determined.
我们对 14 个非手性咪唑衍生物和 5 个手性咪唑衍生物进行了体外测试，作为电鳗乙酰胆碱酯酶酶制剂水解乙酰胆碱的抑制剂。在 25°C、pH8（磷酸盐缓冲液）、离子强度 0.11M 和酶制剂催化活性 0.14U/ml 的反应混合物的分批搅拌反应器中进行了实验。通过原始的 HPLC 方法确定了乙酰胆碱、胆碱和乙酸的实际浓度随时间的变化。对于所有使用的抑制剂，这些时间依赖性与竞争不可逆抑制模型的概率超过 90%相符。我们确定了所有动力学常数，包括定义抑制速率的 k(3)（0.38-5.3M(-1)s(-1)）和对反应混合物中绝对乙酰胆碱酯酶浓度的定量估计（40-110 nM）。
