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Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production
The natural concentration of the anticancer drug Taxol is about 0.02% in yew trees, whereas that of its analogue 7-β-xylosyl-10-deacetyltaxol is up to 0.5%. While this compound is not an intermediate in Taxol biosynthetic route, it can be converted into Taxol by de-glycosylation and acetylation. Her...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5454391/ https://www.ncbi.nlm.nih.gov/pubmed/28516951 http://dx.doi.org/10.1038/ncomms15544 |
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author | Li, Bing-Juan Wang, Hao Gong, Ting Chen, Jing-Jing Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping |
author_facet | Li, Bing-Juan Wang, Hao Gong, Ting Chen, Jing-Jing Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping |
author_sort | Li, Bing-Juan |
collection | PubMed |
description | The natural concentration of the anticancer drug Taxol is about 0.02% in yew trees, whereas that of its analogue 7-β-xylosyl-10-deacetyltaxol is up to 0.5%. While this compound is not an intermediate in Taxol biosynthetic route, it can be converted into Taxol by de-glycosylation and acetylation. Here, we improve the catalytic efficiency of 10-deacetylbaccatin III-10-O-acetyltransferase (DBAT) of Taxus towards 10-deacetyltaxol, a de-glycosylated derivative of 7-β-xylosyl-10-deacetyltaxol to generate Taxol using mutagenesis. We generate a three-dimensional structure of DBAT and identify its active site using alanine scanning and design a double DBAT mutant (DBAT(G38R/F301V)) with a catalytic efficiency approximately six times higher than that of the wild-type. We combine this mutant with a β-xylosidase to obtain an in vitro one-pot conversion of 7-β-xylosyl-10-deacetyltaxol to Taxol yielding 0.64 mg ml(−1) Taxol in 50 ml at 15 h. This approach represents a promising environmentally friendly alternative for Taxol production from an abundant analogue. |
format | Online Article Text |
id | pubmed-5454391 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-54543912017-06-07 Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production Li, Bing-Juan Wang, Hao Gong, Ting Chen, Jing-Jing Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping Nat Commun Article The natural concentration of the anticancer drug Taxol is about 0.02% in yew trees, whereas that of its analogue 7-β-xylosyl-10-deacetyltaxol is up to 0.5%. While this compound is not an intermediate in Taxol biosynthetic route, it can be converted into Taxol by de-glycosylation and acetylation. Here, we improve the catalytic efficiency of 10-deacetylbaccatin III-10-O-acetyltransferase (DBAT) of Taxus towards 10-deacetyltaxol, a de-glycosylated derivative of 7-β-xylosyl-10-deacetyltaxol to generate Taxol using mutagenesis. We generate a three-dimensional structure of DBAT and identify its active site using alanine scanning and design a double DBAT mutant (DBAT(G38R/F301V)) with a catalytic efficiency approximately six times higher than that of the wild-type. We combine this mutant with a β-xylosidase to obtain an in vitro one-pot conversion of 7-β-xylosyl-10-deacetyltaxol to Taxol yielding 0.64 mg ml(−1) Taxol in 50 ml at 15 h. This approach represents a promising environmentally friendly alternative for Taxol production from an abundant analogue. Nature Publishing Group 2017-05-18 /pmc/articles/PMC5454391/ /pubmed/28516951 http://dx.doi.org/10.1038/ncomms15544 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Li, Bing-Juan Wang, Hao Gong, Ting Chen, Jing-Jing Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production |
title | Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production |
title_full | Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production |
title_fullStr | Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production |
title_full_unstemmed | Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production |
title_short | Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production |
title_sort | improving 10-deacetylbaccatin iii-10-β-o-acetyltransferase catalytic fitness for taxol production |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5454391/ https://www.ncbi.nlm.nih.gov/pubmed/28516951 http://dx.doi.org/10.1038/ncomms15544 |
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