Overexpression of BAPT and DBTNBT genes in Taxus baccata in vitro cultures to enhance the biotechnological production of paclitaxel

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• dc.contributor.author Pérez Matas, Edgar
• dc.contributor.author Hidalgo Martínez, Diego
• dc.contributor.author Moyano Claramunt, Elisabet
• dc.contributor.author Palazón Barandela, Javier
• dc.contributor.author Bonfill Baldrich, Ma. Mercedes
• dc.date.accessioned 2023-12-05T07:10:36Z
• dc.date.available 2023-12-05T07:10:36Z
• dc.date.issued 2024
• dc.description.abstract Paclitaxel is one of the most effective anticancer drugs ever developed. Although the most sustainable approach to its production is provided by plant cell cultures, the yield is limited by bottleneck enzymes in the taxane biosynthetic pathway: baccatin-aminophenylpropanoyl-13-O-transferase (BAPT) and 3'-N-debenzoyltaxol N-benzoyltransferase (DBTNBT). With the aim of enhancing paclitaxel production by overcoming this bottleneck, we obtained distinct lines of Taxus baccata in vitro roots, each independently overexpressing either of the two flux-limiting genes, BAPT or DBTNBT, through a Rhizobium rhizogenes A4-mediated transformation. Due to the slow growth rate of the transgenic Taxus roots, they were dedifferentiated to obtain callus lines and establish cell suspensions. The transgenic cells were cultured in a two-stage system and stimulated for taxane production by a dual elicitation treatment with 1 μm coronatine plus 50 mm of randomly methylated-β-cyclodextrins. A high overexpression of BAPT (59.72-fold higher at 48 h) and DBTNBT (61.93-fold higher at 72 h) genes was observed in the transgenic cell cultures, as well as an improved taxane production. Compared to the wild type line (71.01 mg/L), the DBTNBT line produced more than four times higher amounts of paclitaxel (310 mg/L), while the content of this taxane was almost doubled in the BAPT line (135 mg/L). A transcriptional profiling of taxane biosynthetic genes revealed that GGPPS, TXS and DBAT genes were the most reactive to DBTNBT overexpression and the dual elicitation, their expression increasing gradually and constantly. The same genes exhibited a pattern of isolated peaks of expression in the elicited BAPT-overexpressing line.
• dc.description.sponsorship This research was funded by Agencia Estatal de Investigación. REF: PID2020-113438RBI00/AEI/10.13039/501100011033 and by the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) del Departament de Recerca i Universitats de la Generalitat de Catalunya 2021 SGR00693. D.H.-M. is a Postdoctoral researcher at María Zambrano at the University of Barcelona. His contract is financed by the Ministry of Universities, the European Union Next GenerationEU/PRTR.i and the Recovery, Transformation and Resilience Plan.
• dc.format.mimetype application/pdf
• dc.identifier.citation Perez-Matas E, Hidalgo-Martinez D, Moyano E, Palazon J, Bonfill M. Overexpression of BAPT and DBTNBT genes in Taxus baccata in vitro cultures to enhance the biotechnological production of paclitaxel. Plant Biotechnol J. 2024 Jan;22(1):233-47. DOI: 10.1111/pbi.14182
• dc.identifier.doi http://dx.doi.org/10.1111/pbi.14182
• dc.identifier.issn 1467-7644
• dc.identifier.uri http://hdl.handle.net/10230/58439
• dc.language.iso eng
• dc.publisher Wiley
• dc.relation.ispartof Plant Biotechnol J. 2024 Jan;22(1):233-47
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PID2020-113438RBI00
• dc.rights © 2023 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/4.0/
• dc.subject.keyword Taxus baccata transgenic roots
• dc.subject.keyword Bottleneck removal
• dc.subject.keyword Dual elicitation
• dc.subject.keyword Gene overexpression
• dc.subject.keyword Metabolic engineering
• dc.title Overexpression of BAPT and DBTNBT genes in Taxus baccata in vitro cultures to enhance the biotechnological production of paclitaxel
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion