Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Kyoto-Daigaku-Katsura, Nishikyo-ku, Kyoto, Japan.
Macromol Biosci. 2010 Oct 8;10(10):1265-72. doi: 10.1002/mabi.201000189.
A genetically modified His-tagged endoglucanase, EGII(core2), with two active sites in series was immobilized on gold via three different kinds of anchor molecules, and its hydrolytic activity was studied. Immobilization of EGII(core2) was influenced by the chain length and hydrophilicity of anchor molecules. The hydrolytic activity of the immobilized EGII(core2) was nearly the same on either anchor molecule. Interestingly, the immobilized EGII(core2) apparently retained the inherent hydrolytic activity similarly to free EGII(core2). It is therefore considered that the local high concentration of EGII(core2) on the surface should promote the successive hydrolysis of the transient products to show the high hydrolytic activity despite of immobilization.
一种经过基因改造的、带有 His 标签的内切葡聚糖酶 EGII(core2)，具有两个串联的活性位点，通过三种不同的锚定分子固定在金上，并研究了其水解活性。EGII(core2)的固定化受到锚定分子的链长和亲水性的影响。固定化 EGII(core2)在任何一种锚定分子上的水解活性几乎相同。有趣的是，固定化 EGII(core2)显然保留了与游离 EGII(core2)相似的固有水解活性。因此，可以认为表面上 EGII(core2)的局部高浓度应该促进瞬态产物的连续水解，从而表现出高水解活性，尽管存在固定化。
