LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production

Electronic Journal of Biotechnology

Zhao-Yong Yang¹  Ping Zhu²  Tian-Jiao Chen²  Dan-Yang Li³  Sen Zou⁴  Shuai Fan⁵ 

[1] NHC Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China;Research Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052 Henan, China;;Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &;State Key Laboratory of Bioactive Substance and Function of Natural Medicines &

关键词: 10-Deacetyltaxol;    7-β-Xylosyltaxane glycoside hydrolases;    Enzyme kinetics;    Glycoside hydrolase;    Immobilization;    Industrial production;   

DOI  :  

来源: DOAJ

【 摘 要 】

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 Km 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 kcat/Km 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

【 授权许可】

Unknown