Tümtürk H, Karaca N, Demirel G, Sahin F
Gazi University, Faculty of Art and Science, Department of Chemistry, Biotechnology Research Laboratory, 06500 Besevler, Ankara, Turkey.
Int J Biol Macromol. 2007 Feb 20;40(3):281-5. doi: 10.1016/j.ijbiomac.2006.07.004. Epub 2006 Jul 21.
In the present of this study, two novel polymeric matrixes that are poly(N,N-dimethylacrylamide-co-acrylamide) and poly(N-isopropylacrylamide-co-acrylamide)/kappa-Carrageenan was synthesized and applied for immobilization of lipase. For the immobilization of enzyme, two different immobilization procedures have been carried out via covalently binding and entrapment methods. On the free and immobilized enzymes activities, optimum pH, temperature, storage and thermal stability was investigated. The optimum temperature for free, covalently immobilized and entrapped enzymes was found to be 30, 35 and 30 degrees C, respectively. Optimum pH for both free and immobilized enzymes was also observed at pH 8. Maximum reaction rate (Vmax) and Michaelis-Menten constant (Km) were determined for free and immobilized lipases. Furthermore, the reuse numbers of immobilized enzymes also studied. It was observed that after 40th use in 5 days, the retained activities for covalently immobilized and entrapped lipases were found as 39% and 22%, respectively. Storage and thermal stability of enzyme was also increased by as a result of immobilization procedures.
在本研究中,合成了两种新型聚合物基质,即聚(N,N - 二甲基丙烯酰胺 - 共 - 丙烯酰胺)和聚(N - 异丙基丙烯酰胺 - 共 - 丙烯酰胺)/κ-卡拉胶,并将其用于固定化脂肪酶。为了固定化酶,通过共价结合和包埋方法进行了两种不同的固定化程序。研究了游离酶和固定化酶的活性、最佳pH值、温度、储存稳定性和热稳定性。发现游离酶、共价固定化酶和包埋酶的最佳温度分别为30、35和30摄氏度。游离酶和固定化酶的最佳pH值也在pH 8时观察到。测定了游离脂肪酶和固定化脂肪酶的最大反应速率(Vmax)和米氏常数(Km)。此外,还研究了固定化酶的重复使用次数。观察到在5天内第40次使用后,共价固定化脂肪酶和包埋脂肪酶的保留活性分别为39%和22%。固定化程序还提高了酶的储存稳定性和热稳定性。