Background: LXYL-P1-2 is the first reported glycoside hydrolase that can catalyze the transformation of 7-β-xylosyl-10-deacetyltaxol (XDT) to 10-deacetyltaxol (DT) by removing the d-xylosyl group at the C-7 position. Successful synthesis of paclitaxel by one-pot method combining the LXYL-P1-2 and 10-deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) using XDT as a precursor, making LXYL-P1-2 a highly promising enzyme for the industrial production of paclitaxel. The aim of this study was to investigate the catalytic potential of LXYL-P1-2 stabilized on magnetic nanoparticles, the surface of which was modified by Ni²⁺-immobilized cross-linked Fe₃O₄@Histidine.

Results: The diameter of matrix was 20–40 nm. The K_m value of the immobilized LXYL-P1-2 catalyzing XDT (0.145 mM) was lower than that of the free enzyme (0.452 mM), and the k_cat/K_m value of immobilized enzyme (12.952 mM s⁻¹) was higher than the free form (8.622 mM s⁻¹). The immobilized form maintained 50% of its original activity after 15 cycles of reuse. In addition, the stability of immobilized LXYL-P1-2, maintained 84.67% of its initial activity, improved in comparison with free form after 30 d storage at 4°C.

Conclusions: This investigation not only provides an effective procedure for biocatalytic production of DT, but also gives an insight into the application of magnetic material immobilization technology.