Topchieva I N, Sorokina E M, Kurganov B I, Zhulin V M, Makarova Z G
Biokhimiia. 1996 Jun;61(6):1041-5.
A new method of formation of non-covalent adducts based on an amphiphilic diblock copolymer of ethylene and propylene oxides with molecular mass of 2 kDa and alpha-chymotrypsin (ChT) under high pressure, has been developed. The composition of the complexes corresponds to seven polymer molecules per one ChT molecule in the pressure range of 1.1 to 400 MPa. The complexes fully retain the catalytic activity. Kinetic constants (Km and kcat) for enzymatic hydrolysis of N-benzoyl-L-tyrosine ethyl ester catalyzed by the complexes are identical with the corresponding values for native ChT. Analysis of kinetics of thermal inactivation of the complexes revealed that the constant of the rate of the slow inactivation step is markedly lower than for ChT.
已开发出一种基于分子量为2 kDa的环氧乙烷和环氧丙烷两亲性二嵌段共聚物与α-胰凝乳蛋白酶(ChT)在高压下形成非共价加合物的新方法。在1.1至400 MPa的压力范围内,复合物的组成对应于每一个ChT分子有七个聚合物分子。这些复合物完全保留了催化活性。复合物催化N-苯甲酰-L-酪氨酸乙酯酶促水解的动力学常数(Km和kcat)与天然ChT的相应值相同。对复合物热失活动力学的分析表明,缓慢失活步骤的速率常数明显低于ChT的。