Biosynthesis of novel anticoagulant substances, α-salicin and α-isosalicin, using sucrose phosphorylase.

This study reports the enzymatic synthesis of novel anticoagulant glycosides, α-salicin and α-isosalicin, through the transglycosylation of o-hydroxybenzyl alcohol (salicyl alcohol) catalyzed by marine-derived sucrose phosphorylase Suc75290. Under optimized conditions (240 g/L sucrose, 12 g/L o-hydroxybenzyl alcohol, 150 U/mL enzyme, pH 7.5, 45 °C, 20 h), a conversion rate of 79.17 ± 0.64 % was achieved, yielding 8.57 ± 0.33 g/L α-salicin and 0.52 ± 0.19 g/L α-isosalicin. Structural characterization by NMR confirmed the α-configuration of both compounds. In vitro anticoagulant assays demonstrated that these glycosides significantly prolonged activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT), while reducing fibrinogen (FIB) levels, indicating dual inhibition of intrinsic and extrinsic coagulation pathways. Molecular docking revealed key interactions between salicyl alcohol and Suc75290's active site. This work establishes an efficient enzymatic route to α-anomeric salicin analogs with therapeutic potential, circumventing the limitations of chemical synthesis. These two novel anticoagulant glycoside compounds enzymatically synthesized provide new research ideas for the development of anticoagulant drugs.