College of Food Science and Engineering, Northwest University, 229 North Taibai Road, Xi'an, 710000, Shaanxi, China.
College of Food Science and Engineering, Northwest University, 229 North Taibai Road, Xi'an, 710000, Shaanxi, China.
J Biotechnol. 2018 Sep 10;281:67-73. doi: 10.1016/j.jbiotec.2018.06.343. Epub 2018 Jun 19.
Phospholipase D (PLD) was firstly hyperactivated by the bio-imprinting and, then, these hyperactivated PLD molecules were immobilized by adsorption and precipitation, followed by cross-linking. The high degree of conformational rigidification provided by intra- and intermolecular cross-linking can make PLD "remember" imprint-induced characteristics even in aqueous solutions. The obtained immobilized PLD showed the excellent catalytic performance. The maximum activity of immobilized PLD reached 166953 U/g, which was approximately 14 times higher than that of free form (11922 U/g). Moreover, the selectivity of PLD was significantly enhanced after immobilization. The yield of PG and PA reached 94.0% and 5.96%, respectively. Compared with the serious hydrolysis in the free PLD (35.3% yield of PA), the side reaction was minimized in this work. This may be the first description of the remarkably high improvement of the activity and selectivity of PLD through the immobilization technology.
磷脂酶 D（PLD）首先被生物印迹强烈激活，然后这些被强烈激活的 PLD 分子通过吸附和沉淀固定，接着进行交联。分子内和分子间交联提供的高度构象僵化可以使 PLD“记住”印迹诱导的特征，即使在水溶液中也是如此。所得到的固定化 PLD 表现出优异的催化性能。固定化 PLD 的最大活性达到 166953 U/g，约为游离形式（11922 U/g）的 14 倍。此外，固定化后 PLD 的选择性显著提高。PG 和 PA 的产率分别达到 94.0%和 5.96%。与游离 PLD 中严重的水解（PA 的产率为 35.3%）相比，本工作中最小化了副反应。这可能是首次通过固定化技术显著提高 PLD 的活性和选择性的描述。
