Cao Rong, Gu Zhenyu, Patterson Gary D, Armitage Bruce A
Department of Chemistry, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, USA.
J Am Chem Soc. 2004 Jan 28;126(3):726-7. doi: 10.1021/ja039502d.
The enzyme beta-galactosidase has been immobilized through incorporation into a selectively soluble microgel, prepared from DNA, biotinylated peptide nucleic acid (PNA), and the protein avidin. The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficient hydrolysis of a small-molecule substrate occurred at 37 degrees , but cooling and centrifuging led to precipitation of the microgels and product separation. The microgels were then reconstituted by adding fresh buffer and shaking. The enzymatic activity was completely recovered through repeated cycles. This method should be generalizable to a wide variety of other enzymes and substrates.
β-半乳糖苷酶已通过掺入一种选择性可溶的微凝胶中实现固定化，该微凝胶由DNA、生物素化肽核酸（PNA）和抗生物素蛋白制备而成。该酶与抗生物素蛋白结合，使其能够直接整合到微凝胶网络中。小分子底物在37摄氏度时发生高效水解，但冷却和离心导致微凝胶沉淀并实现产物分离。然后通过加入新鲜缓冲液并振荡来重构微凝胶。通过重复循环，酶活性完全恢复。该方法应可推广到多种其他酶和底物。
