Ivanova Tz, Mircheva K, Dobreva G, Panaiotov I, Proust J E, Verger R
Biophysical Chemistry Laboratory, Department of Physical Chemistry, University of Sofia, J. Bourchier Street 1, 1126 Sofia, Bulgaria.
Colloids Surf B Biointerfaces. 2008 May 1;63(1):91-100. doi: 10.1016/j.colsurfb.2007.11.004. Epub 2007 Nov 21.
The hydrolysis catalyzed by Humicola lanuginosa lipase (HLL) of pure tricaprylin (TC) or stearate of polyethylene glycol 1500 (PEG-St) as well as their mixtures spread as monomolecular films were studied. The catalytic transformation of the two substrates TC or PEG-St into their respective reaction products was detected by measuring simultaneously the decrease in the film area and the surface potential using the "zero order" trough at constant surface pressure. A kinetic model describing the enzymatic hydrolysis was developed. The surface concentrations of the two substrates and their respective reaction products as well as the values of the global kinetic constants of hydrolysis were determined. The experimentally obtained global kinetic constants of the catalytic action of HLL against TC and PEG-St present in mixed monolayers of TC/PEG-St are approximately the same as in the case of pure monolayers. These obtained results give some indications that the activity of enzyme is not significantly affected by the different molecular environments in the mixed monolayers.
研究了由疏棉状嗜热丝孢菌脂肪酶(HLL)催化的纯三辛酸甘油酯(TC)或聚乙二醇1500硬脂酸酯(PEG-St)及其混合物铺展成单分子膜时的水解反应。通过在恒定表面压力下使用“零级”槽同时测量膜面积的减小和表面电势,检测了两种底物TC或PEG-St向其各自反应产物的催化转化。建立了描述酶促水解的动力学模型。测定了两种底物及其各自反应产物的表面浓度以及水解的整体动力学常数的值。实验获得的HLL对存在于TC/PEG-St混合单分子层中的TC和PEG-St的催化作用的整体动力学常数与纯单分子层的情况大致相同。这些结果表明,混合单分子层中不同的分子环境对酶的活性没有显著影响。