Improving phospholipase D activity and selectivity by bio-imprinting-immobilization to produce phosphatidylglycerol.

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/gprotein, which was approximately 14 times higher than that of free form (11922 U/gprotein). 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.