Institute of Technical Biocatalysis, Hamburg University of Technology, Denickestr. 15, 21073 Hamburg, Germany.
J Biotechnol. 2013 Aug 10;167(1):1-7. doi: 10.1016/j.jbiotec.2013.06.002. Epub 2013 Jun 14.
This study shows the effect of site-directed enzyme immobilization upon the enzyme activity of covalently bound glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Immobilization points were introduced at sterically accessible sites in order to control the protein's orientation and twice as much activity was recovered in comparison to conventionally immobilized enzyme. Immobilization of G6PDH via genetically engineered cysteine provided a simple, but effective method to control the immobilization process. G6PDH variants with cysteine close to the active center (L218C), close to the dimer interface (D205C) as well as far from the active center (D453C) showed changes in activity and the efficacy of immobilization.
本研究展示了定向酶固定化对来自肠膜明串珠菌的共价结合葡萄糖-6-磷酸脱氢酶的酶活性的影响。固定化点位于空间可及的位点，以控制蛋白质的方向，与传统固定化酶相比，活性回收增加了一倍。通过基因工程半胱氨酸固定化 G6PDH 提供了一种简单而有效的方法来控制固定化过程。靠近活性中心（L218C）、靠近二聚体界面（D205C）以及远离活性中心（D453C）的半胱氨酸的 G6PDH 变体显示出活性和固定化效果的变化。
