LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production
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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-03-01
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| Series: | Electronic Journal of Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345820300658 |
| _version_ | 1818870412827688960 |
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| author | Sen Zou Tian-Jiao Chen Dan-Yang Li Shuai Fan Zhao-Yong Yang Ping Zhu |
| author_facet | Sen Zou Tian-Jiao Chen Dan-Yang Li Shuai Fan Zhao-Yong Yang Ping Zhu |
| author_sort | Sen Zou |
| collection | DOAJ |
| description | 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 |
| first_indexed | 2024-12-19T12:06:37Z |
| format | Article |
| id | doaj.art-b178defd09da4b64ad1095b8fe6bd8eb |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-12-19T12:06:37Z |
| publishDate | 2021-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-b178defd09da4b64ad1095b8fe6bd8eb2022-12-21T20:22:20ZengElsevierElectronic Journal of Biotechnology0717-34582021-03-01501015LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel productionSen Zou0Tian-Jiao Chen1Dan-Yang Li2Shuai Fan3Zhao-Yong Yang4Ping Zhu5Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China; Research Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052 Henan, ChinaState Key Laboratory of Bioactive Substance and Function of Natural Medicines & NHC Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaKey Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaKey Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, ChinaKey Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China; Corresponding authors.State Key Laboratory of Bioactive Substance and Function of Natural Medicines & NHC Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China; Corresponding authors.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.2020.12.005http://www.sciencedirect.com/science/article/pii/S071734582030065810-Deacetyltaxol7-β-Xylosyltaxane glycoside hydrolasesEnzyme kineticsGlycoside hydrolaseImmobilizationIndustrial production |
| spellingShingle | Sen Zou Tian-Jiao Chen Dan-Yang Li Shuai Fan Zhao-Yong Yang Ping Zhu LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production Electronic Journal of Biotechnology 10-Deacetyltaxol 7-β-Xylosyltaxane glycoside hydrolases Enzyme kinetics Glycoside hydrolase Immobilization Industrial production |
| title | LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production |
| title_full | LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production |
| title_fullStr | LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production |
| title_full_unstemmed | LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production |
| title_short | LXYL-P1-2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production |
| title_sort | lxyl p1 2 immobilized on magnetic nanoparticles and its potential application in paclitaxel production |
| topic | 10-Deacetyltaxol 7-β-Xylosyltaxane glycoside hydrolases Enzyme kinetics Glycoside hydrolase Immobilization Industrial production |
| url | http://www.sciencedirect.com/science/article/pii/S0717345820300658 |
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