Bacheva A V, Baibak O V, Belyaeva A V, Oksenoit E S, Velichko T I, Lysogorskaya E N, Gladilin A K, Lozinsky V I, Filippova I Yu
Faculty of Chemistry, Lomonosov Moscow State University, Moscow, 119992, Russia.
Biochemistry (Mosc). 2003 Nov;68(11):1261-6. doi: 10.1023/b:biry.0000009142.88974.85.
The activity and stability of native subtilisin 72, its complex with poly(acrylic acid), and subtilisin covalently attached to poly(vinyl alcohol) cryogel were studied in aqueous and organic media by hydrolysis of specific chromogenic peptide substrates. Kinetic parameters of the hydrolysis of Glp-Ala-Ala-Leu-pNA by native subtilisin and its complex with poly(acrylic acid) were determined. Based on the comparative study of stability of native and modified subtilisins in media of various compositions, it was established that covalent immobilization of subtilisin on poly(vinyl alcohol) cryogel is the most effective approach to improve enzyme stability in water as well as in mixtures with low water content.
通过特定显色肽底物的水解反应，在水性和有机介质中研究了天然枯草杆菌蛋白酶72、其与聚丙烯酸的复合物以及共价连接到聚乙烯醇冷冻凝胶上的枯草杆菌蛋白酶的活性和稳定性。测定了天然枯草杆菌蛋白酶及其与聚丙烯酸的复合物水解Glp-Ala-Ala-Leu-pNA的动力学参数。基于对天然和修饰枯草杆菌蛋白酶在各种组成介质中稳定性的比较研究，确定将枯草杆菌蛋白酶共价固定在聚乙烯醇冷冻凝胶上是提高酶在水中以及与低含水量混合物中稳定性的最有效方法。
