eprintid: 10200105 rev_number: 9 eprint_status: archive userid: 699 dir: disk0/10/20/01/05 datestamp: 2024-11-14 09:44:43 lastmod: 2024-11-14 09:44:43 status_changed: 2024-11-14 09:44:43 type: article metadata_visibility: show sword_depositor: 699 creators_name: Escrich, A creators_name: Jonguitud-Borrego, N creators_name: Malci, K creators_name: Sanchez-Muñoz, R creators_name: Palazon, J creators_name: Rios-Solis, L creators_name: Moyano, E title: A novel step towards the heterologous biosynthesis of paclitaxel: Characterization of T1βOH taxane hydroxylase ispublished: pub divisions: UCL divisions: B04 divisions: F47 keywords: Enzyme characterization, Heterologous production, Paclitaxel, T1βOH, Taxane hydroxylase, Yeast consortia, Paclitaxel, Saccharomyces cerevisiae, Mixed Function Oxygenases, Molecular Docking Simulation, Metabolic Engineering, Taxoids, Bridged-Ring Compounds note: © 2024 The Authors. Published by Elsevier Inc. on behalf of International Metabolic Engineering Society. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). abstract: In the quest for innovative cancer therapeutics, paclitaxel remains a cornerstone in clinical oncology. However, its complex biosynthetic pathway, particularly the intricate oxygenation steps, has remained a puzzle in the decades following the characterization of the last taxane hydroxylase. The high divergence and promiscuity of enzymes involved have posed significant challenges. In this study, we adopted an innovative approach, combining in silico methods and functional gene analysis, to shed light on this elusive pathway. Our molecular docking investigations using a library of potential ligands uncovered TB574 as a potential missing enzyme in the paclitaxel biosynthetic pathway, demonstrating auspicious interactions. Complementary in vivo assays utilizing engineered S. cerevisiae strains as novel microbial cell factory consortia not only validated TB574's critical role in forging the elusive paclitaxel intermediate, T5αAc-1β,10β-diol, but also achieved the biosynthesis of paclitaxel precursors at an unprecedented yield including T5αAc-1β,10β-diol with approximately 40 mg/L. This achievement is highly promising, offering a new direction for further exploration of a novel metabolic engineering approaches using microbial consortia. In conclusion, our study not only furthers study the roles of previously uncharacterized enzymes in paclitaxel biosynthesis but also forges a path for pioneering advancements in the complete understanding of paclitaxel biosynthesis and its heterologous production. The characterization of T1βOH underscores a significant leap forward for future advancements in paclitaxel production using heterologous systems to improve cancer treatment and pharmaceutical production, thereby holding immense promise for enhancing the efficacy of cancer therapies and the efficiency of pharmaceutical manufacturing. date: 2024-09-01 date_type: published publisher: Elsevier BV official_url: http://dx.doi.org/10.1016/j.ymben.2024.08.005 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2312686 doi: 10.1016/j.ymben.2024.08.005 medium: Print-Electronic pii: S1096-7176(24)00108-3 lyricists_name: Rios-Solis, Leonardo lyricists_id: LRIOS73 actors_name: Rios-Solis, Leonardo actors_id: LRIOS73 actors_role: owner funding_acknowledgements: [Royal Society of Medicine]; [CONACYT]; [Agencia Estatal de Investigación]; [Engineering and Physical Sciences Research Council]; [Republic of Turkey Ministry of National Education]; [Universitat Pompeu Fabra]; [British Council] full_text_status: public publication: Metabolic Engineering volume: 85 pagerange: 201-212 event_location: Belgium issn: 1096-7176 citation: Escrich, A; Jonguitud-Borrego, N; Malci, K; Sanchez-Muñoz, R; Palazon, J; Rios-Solis, L; Moyano, E; (2024) A novel step towards the heterologous biosynthesis of paclitaxel: Characterization of T1βOH taxane hydroxylase. Metabolic Engineering , 85 pp. 201-212. 10.1016/j.ymben.2024.08.005 <https://doi.org/10.1016/j.ymben.2024.08.005>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10200105/1/Rios-Solis_1-s2.0-S1096717624001083-main.pdf