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.

How to cite

Zou S, Chen TJ, Li DY, et al. LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production. Electron J Biotechnol 2021;50.https://doi.org/10.1016/j.ejbt.2020.12.005

中文翻译：

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固定在磁性纳米粒子上的LXYL-P1-2及其在紫杉醇生产中的潜在应用

背景

LXYL-P1-2是第一个报道的糖苷水解酶，可通过去除C-7位的d-木糖基来催化7-β-木糖基-10-去乙酰基紫杉醇（XDT）向10-去乙酰基紫杉醇（DT）的转化。使用XDT作为前体，通过一锅法成功结合LXYL-P1-2和10-去乙酰基浆果赤霉素III-10-β- O-乙酰基转移酶（DBAT）合成紫杉醇，使LXYL-P1-2成为用于紫杉醇的极有前途的酶紫杉醇的工业生产。这项研究的目的是研究稳定在磁性纳米粒子上的LXYL-P1-2的催化潜力，该粒子的表面被Ni ²⁺固定的交联Fe ₃ O ₄ @组氨酸修饰。

结果

基质的直径为20–40 nm。固定化LXYL-P1-2催化XDT的K _m值（0.145 mM）低于游离酶的K _m值（0.452 mM），固定化酶的k _cat / K _m值（12.952 mM s ^-1）为高于自由形式（8.622 mM s ^-1）。在重复使用15个循环后，固定化形式保持其原始活性的50％。此外，固定化的LXYL-P1-2的稳定性保持了其初始活性的84.67％，与在4°C下储存30天后的游离形式相比，稳定性有所提高。

结论

这项研究不仅为DT的生物催化生产提供了有效的程序，而且还为磁性材料固定技术的应用提供了见识。

如何引用

邹S，陈天杰，李迪，等。LXYL-P1-2固定在磁性纳米粒子上及其在紫杉醇生产中的潜在应用。电子生物技术杂志2021; 50.https：//doi.org/10.1016/j.ejbt.2020.12.005