One-pot synthesis and biochemical characterization of the novel magnetic phospholipase D nanoflowers modified with polyphenols

Phospholipase D (PLD) can catalyze the hydrolytic cleavage of phosphodiester bonds and transphosphatidylation and has been widely employed in the chemical and pharmaceutical fields. To improve the stability and reusability of PLD, an immobilized enzyme system was developed using magnetic nanoparticles modified with polyphenols (dopamine and tannic acid) and metal hybrid nanoflowers to co-immobilize phospholipase D (PLD). This immobilization technique aimed to improve the enzymatic performance through increased surface area, reduced mass transfer limitations, and facilitated reuse. The hydrolytic and transphosphatidylation activities of PLD immobilized on polyphenol-modified magnetic nanoflowers (PDA-MNFs and TA-MNFs) were evaluated and compared to free PLD. PDA-MNFs and TA-MNFs showed significantly enhanced hydrolytic activity (19.9-fold and 21.7-fold increases, respectively) and transphosphatidylation activity (over 1.79-fold and 1.96-fold increases, respectively) compared to free PLD. Temperature and pH stability were also markedly improved for the immobilized PLD. After 10 reuse cycles, PDA-MNFs and TA-MNFs retained over 65 % of their initial transphosphatidylation activity. Therefore, this co-deposition and polymerization immobilization strategy using polyphenol-modified magnetic nanoflowers has the potential for efficiently increasing enzymatic performance and synthesizing industrially relevant compounds like phosphatidic acid and phosphatidylserine