A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology
Engineered plant cell lines have the potential to achieve enhanced metabolite production rates, providing a high-yielding source of compounds of interest. Improving the production of taxanes, pharmacologically valuable secondary metabolites of Taxus spp., is hindered by an incomplete knowledge of th...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-05-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fbioe.2020.00410/full |
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| author | Raul Sanchez-Muñoz Edgar Perez-Mata Lorena Almagro Rosa M. Cusido Mercedes Bonfill Javier Palazon Elisabeth Moyano |
| author_facet | Raul Sanchez-Muñoz Edgar Perez-Mata Lorena Almagro Rosa M. Cusido Mercedes Bonfill Javier Palazon Elisabeth Moyano |
| author_sort | Raul Sanchez-Muñoz |
| collection | DOAJ |
| description | Engineered plant cell lines have the potential to achieve enhanced metabolite production rates, providing a high-yielding source of compounds of interest. Improving the production of taxanes, pharmacologically valuable secondary metabolites of Taxus spp., is hindered by an incomplete knowledge of the taxane biosynthetic pathway. Of the five unknown steps, three are thought to involve cytochrome P450-like hydroxylases. In the current work, after an in-depth in silico characterization of four candidate enzymes proposed in a previous cDNA-AFLP assay, TB506 was selected as a candidate for the hydroxylation of the taxane side chain. A docking assay indicated TB506 is active after the attachment of the side chain based on its affinity to the ligand 3′N-dehydroxydebenzoyltaxol. Finally, the involvement of TB506 in the last hydroxylation step of the paclitaxel biosynthetic pathway was confirmed by functional assays. The identification of this hydroxylase will contribute to the development of alternative sustainable paclitaxel production systems using synthetic biology techniques. |
| first_indexed | 2024-12-14T12:45:05Z |
| format | Article |
| id | doaj.art-66e86be3ae0d4c358df2d92dea3e4c51 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-14T12:45:05Z |
| publishDate | 2020-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-66e86be3ae0d4c358df2d92dea3e4c512022-12-21T23:00:48ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-05-01810.3389/fbioe.2020.00410538792A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic BiologyRaul Sanchez-Muñoz0Edgar Perez-Mata1Lorena Almagro2Rosa M. Cusido3Mercedes Bonfill4Javier Palazon5Elisabeth Moyano6Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, SpainSecció de Fisiologia Vegetal, Facultat de Farmacia, Universitat de Barcelona, Barcelona, SpainDepartamento de Biología Vegetal, Facultad de Biología, Universidad de Murcia, Murcia, SpainSecció de Fisiologia Vegetal, Facultat de Farmacia, Universitat de Barcelona, Barcelona, SpainSecció de Fisiologia Vegetal, Facultat de Farmacia, Universitat de Barcelona, Barcelona, SpainSecció de Fisiologia Vegetal, Facultat de Farmacia, Universitat de Barcelona, Barcelona, SpainDepartament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, SpainEngineered plant cell lines have the potential to achieve enhanced metabolite production rates, providing a high-yielding source of compounds of interest. Improving the production of taxanes, pharmacologically valuable secondary metabolites of Taxus spp., is hindered by an incomplete knowledge of the taxane biosynthetic pathway. Of the five unknown steps, three are thought to involve cytochrome P450-like hydroxylases. In the current work, after an in-depth in silico characterization of four candidate enzymes proposed in a previous cDNA-AFLP assay, TB506 was selected as a candidate for the hydroxylation of the taxane side chain. A docking assay indicated TB506 is active after the attachment of the side chain based on its affinity to the ligand 3′N-dehydroxydebenzoyltaxol. Finally, the involvement of TB506 in the last hydroxylation step of the paclitaxel biosynthetic pathway was confirmed by functional assays. The identification of this hydroxylase will contribute to the development of alternative sustainable paclitaxel production systems using synthetic biology techniques.https://www.frontiersin.org/article/10.3389/fbioe.2020.00410/fulltaxane hydroxylasepaclitaxelprotoplasts transfectioncytochrome P450biotechnological productionbiotransformation |
| spellingShingle | Raul Sanchez-Muñoz Edgar Perez-Mata Lorena Almagro Rosa M. Cusido Mercedes Bonfill Javier Palazon Elisabeth Moyano A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology Frontiers in Bioengineering and Biotechnology taxane hydroxylase paclitaxel protoplasts transfection cytochrome P450 biotechnological production biotransformation |
| title | A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology |
| title_full | A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology |
| title_fullStr | A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology |
| title_full_unstemmed | A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology |
| title_short | A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology |
| title_sort | novel hydroxylation step in the taxane biosynthetic pathway a new approach to paclitaxel production by synthetic biology |
| topic | taxane hydroxylase paclitaxel protoplasts transfection cytochrome P450 biotechnological production biotransformation |
| url | https://www.frontiersin.org/article/10.3389/fbioe.2020.00410/full |
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