Immobilization of Phospholipase D on Fe₃O₄@SiO₂-Graphene Oxide nanocomposites: A Strategy to Improve Catalytic Stability and Reusability in the Efficient Production of Phosphatidylserine

Phospholipase D (PLD) plays a pivotal role in the biosynthesis of phosphatidylserine (PS), but its practical application is constrained by limitations in stability and reusability. In this study, we successfully fabricated the Fe3O4@SiO2-graphene oxide (GO) nanocomposite by chemical binding of Fe3O4@SiO2 and GO. Subsequently, PLD was immobilized onto the nanocomposite via physical adsorption, with the aim of enhancing catalytic stability, reducing mass transfer resistance, and improving reusability. Under optimal conditions, the immobilization efficiency reached 84.4%, with a PLD loading capacity of 111.4 mg/gsupport. The optimal pH for PS production by immobilized PLD shifted from 6.0 to 6.5, while the optimal temperature increased from 45 °Cto 50 °C. Notably, the immobilized PLD demonstrated a shorter reaction time and a higher PS yield, achieving a 95.4% yield within 90 min, compared to the free PLD (78.1% yield within 150 min), representing a 1.04-fold improvement in production efficiency. Furthermore, the immobilized PLD exhibited outstanding storage stability and thermal stability, along with remarkable reusability. Even after being reused for 10 cycles, the PS yield still stays as high as 78.3%. These findings strongly suggest that the Fe3O4@SiO2-GO immobilized PLD has potential for the efficient production of PS.