Emerging Technologies Empowering the Biosynthesis of Paclitaxel

Abstract

Paclitaxel (Taxol) is a clinically important diterpenoid anticancer drug whose industrial production remains constrained by limited Taxusresources and semi-synthetic routes. Driven by the rapid advancement of genome mining and synthetic biology technologies, the past two years have witnessed substantial breakthroughs in elucidating the biosynthetic pathway of paclitaxel. The pathway constitutes an exceptionally complex biosynthetic network comprising approximately 20 enzymatic steps, predominantly catalyzed by cytochrome P450 monooxygenases, 2-oxoglutarate-dependent dioxygenases (ODDs), and acyltransferases. Nevertheless, microbial production of paclitaxel remains highly obstructed, largely due to inefficient catalytic abilities, enzyme promiscuities, and complex metabolic fluxes. This review summarizes recent progress in elucidating the evolutionary origins and catalytic mechanistic basis of the paclitaxel biosynthetic pathway, with particular emphasis on the emerging technologies and catalytic mechanism studies. Furthermore, current challenges and perspectives for constructing efficient artificial biosynthetic pathways are discussed, providing insights into the future biotechnological production of paclitaxel.

Keywords

Paclitaxel / Pathway analysis / P450 enzyme / Enzyme modification / Synthetic biology

Statement of the Use of Generative AI and AI-Assisted Technologies in the Writing Process

During the preparation of this manuscript, the authors used DeepSeek and ChatGPT to polish specific sentences to better align with English writing standards. Following the use of these tools, the authors carefully reviewed and edited the content as necessary and assume full responsibility for the content of the published article.

Author Contributions

X.L.: designed, wrote and revised the paper. Y.Z.: wrote the paper and collected the data. J.L.: wrote the paper and collected the data. H.J.: revised the manuscript.

Ethics Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data sharing not applicable to this article as no datasets were generated or analysed during the current study.

Funding

This work was supported by the National Natural Science Foundation of China (Grant No. 32371499), Natural Science Foundation of Wuhan (2025040601020158), Science and Technology Major Project of Guangxi (Guike AA24206048, Guike AA24206050-3, Guike JF2504850012), Hubei University of Technology High-Level Talent Research Startup Fund Program (4301/00960), Hubei Provincial Key R&D Major Projects (2025BCA006), Youth Science and Technology Talent Cultivation Special Program of Hubei Province (2025DJA024).

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References

[1] Prota AE, Katja B, Didier Z, Field JJ, José Fernando D, Karl-Heinz A, et al. Molecular mechanism of action of microtubulestabilizing anticancer agents. Science 2013, 339, 587-590. DOI:10.1126/science.1230582

[2] Li C, Wang Y, Yin X, Sun X, Sui S, Liu J, et al. Novel alpha-KG/Fe(II)-Dependent Dioxygenases Catalyzing C1beta-Hydroxylation and Construction of 5/7/6-Skeleton of Highly Oxygenated Taxoids. Angew. Chem. Int. Ed. 2025, 137, e202517041. DOI:10.1002/ange.202517041

[3] Winegar PH, Astolfi MC, Holm SF, Hudson GA, Keasling JD. Advances in the microbial biosynthesis of therapeutic terpenoids. Curr. Opin. Biotechnol. 2025, 96, 103374. DOI:10.1016/j.copbio.2025.103374

[4] Zhu Y, Liang F, Chai P, Kampranis SC, Zhao Y. Recent Advances in the Biosynthesis and Biotechnological Production of Taxol. Engineering 2025, 4, 65-75. DOI:10.1016/j.eng.2025.12.018

[5] Tan CL, Yu X, Feng HC, Gershenzon J, Liu Y, Li SH. A synthetic biology roadmap for sustainable production of the plantoriginated anti-cancer drug paclitaxel. Trends Biotechnol. 2026. DOI:10.1016/j.tibtech.2025.11.013

[6] Xiong X, Gou J, Liao Q, Li Y, Zhou Q, Bi G, et al. The Taxus genome provides insights into paclitaxel biosynthesis. Nat. Plants 2021, 7, 1026-1036. DOI:10.1038/s41477-021-00963-5

[7] Cheng J, Wang X, Liu X, Zhu X, Li Z, Chu H, et al. Chromosome-level genome of Himalayan yew provides insights into the origin and evolution of the paclitaxel biosynthetic pathway. Mol. Plant 2021, 14, 1199-1209. DOI:10.1016/j.molp.2021.04.015

[8] Song C, Fu F, Yang L, Niu Y, Tian Z, He X, et al. Taxus yunnanensis genome offers insights into gymnosperm phylogeny and taxol production. Commun. Biol. 2021, 4, 1203. DOI:10.1038/s42003-021-02697-8

[9] Zhang Y, Scossa F, Fernie AR. The genomes of Taxus species unveil novel candidates in the biosynthesis of taxoids. Mol. Plant 2021, 14, 1773-1775. DOI:10.1016/j.molp.2021.08.017

[10] Wang M, Ma R, Fang Z, Zhang L, Zhang Y, Zheng M, et al. Analysis genome of Pseudotaxus chienii reveals insights into the origin and evolution of taxane biosynthesis. Nat. Commun. 2025, 17, 1088. DOI:10.1038/s41467-025-67849-4

[11] McClune CJ, Liu JC, Wick C, De La Pena R, Lange BM, Fordyce PM, et al. Discovery of FoTO1 and Taxol genes enables biosynthesis of baccatin III. Nature 2025, 643, 582-592. DOI:10.1038/s41586-025-09090-z

[12] Jiang B, Gao L, Wang H, Sun Y, Zhang X, Ke H, et al. Characterization and heterologous reconstitution of Taxus biosynthetic enzymes leading to baccatin III. Science 2024, 383, 622-629. DOI:10.1126/science.adj3484

[13] Yang C, Wang Y, Su Z, Xiong L, Wang P, Lei W, et al. Biosynthesis of the highly oxygenated tetracyclic core skeleton of Taxol. Nat. Commun. 2024, 15, 2339. DOI:10.1038/s41467-024-46583-3

[14] Li C, Yin X, Wang S, Sui S, Liu J, Sun X, et al. A Cytochrome P450 Enzyme Catalyses Oxetane Ring Formation in Paclitaxel Biosynthesis. Angewandte Chem. Int. Ed. 2024, 63, e202407070. DOI:10.1002/anie.202407070

[15] Zhang Y, Wiese L, Fang H, Alseekh S, Perez de Souza L, Scossa F, et al. Synthetic biology identifies the minimal gene set required for Paclitaxel biosynthesis in a plant chassis. Mol. Plant 2023, 16, 1951-1961. DOI:10.1016/j.molp.2023.10.016

[16] Liang F, Xie Y, Zhang C, Zhao Y, Motawia MS, Kampranis SC. Elucidation of the final steps in Taxol biosynthesis and its biotechnological production. Nat. Synth. 2025, 4, 1212-1222. DOI:10.1038/s44160-025-00800-z

[17] Lai M, Weng Y, Wei J, Lu XY, Huang Z, Li H, et al. Designing an oxidase toolbox for site-directed oxidation of taxanes. Nat. Commun. 2025, 17, 841. DOI:10.1038/s41467-025-67544-4

[18] Tang X, Zhang X, Long LP, Wu S, Fan XY, Liu J, et al. Versatile 2-Oxoglutarate-Dependent Dioxygenases Catalyze Radical-Mediated Multifunctional Skeleton Reconstructions and Oxidation Modifications of Taxoids. J. Am. Chem. Soc. 2026, 148, 11333-11343. DOI:10.1021/jacs.6c01169

[19] Bian G, Yuan Y, Tao H, Shi X, Zhong X, Han Y, et al. Production of taxadiene by engineering of mevalonate pathway in Escherichia coli and endophytic fungus Alternaria alternata TPF6. Biotechnol. J. 2017, 12, 1600697. DOI:10.1002/biot.201600697

[20] Williams DC, Wildung MR, Jin AQ, Dalal D, Oliver JS, Coates RM, et al. Heterologous expression and characterization of a “Pseudomature” form of taxadiene synthase involved in paclitaxel (Taxol) biosynthesis and evaluation of a potential intermediate and inhibitors of the multistep diterpene cyclization reaction. Arch. Biochem. Biophys. 2000, 379, 137-146. DOI:10.1006/abbi.2000.1865

[21] Edgar S, Zhou K, Qiao K, King JR, Simpson JH, Stephanopoulos G. Mechanistic Insights into Taxadiene Epoxidation by Taxadiene-5alpha-Hydroxylase. ACS Chem. Biol. 2016, 11, 460-469. DOI:10.1021/acschembio.5b00767

[22] Edgar S, Li FS, Qiao K, Weng JK, Stephanopoulos G. Engineering of Taxadiene Synthase for Improved Selectivity and Yield of a Key Taxol Biosynthetic Intermediate. ACS Synth. Biol. 2017, 6, 201-205. DOI:10.1021/acssynbio.6b00206

[23] Zhao Y, Liang F, Xie Y, Duan Y-T, Andeadelli A, Pateraki I, et al. Oxetane Ring Formation in Taxol Biosynthesis Is Catalyzed by a Bifunctional Cytochrome P450 Enzyme. J. Am. Chem. Soc. 2023, 146, 801-810. DOI:10.1021/jacs.3c10864

[24] Li C, Yin X, Dai J. The History of Studies on Oxetane Ring Formation in Paclitaxel Biosynthesis. Chembiochem 2025, 26, e202400947. DOI:10.1002/cbic.202400947

[25] Tong Y, Luo YF, Gao W. Biosynthesis of paclitaxel using synthetic biology. Phytochem. Rev. 2021, 21, 863-877. DOI:10.1007/s11101-021-09766-0

[26] Liu JCT, De La Pena R, Tocol C, Sattely ES. Reconstitution of early paclitaxel biosynthetic network. Nat. Commun. 2024, 15, 1419. DOI:10.1038/s41467-024-45574-8

[27] Li C, Sun X, Chen R, Xie K, Chen D, Liu J, et al. The discovery of missing taxane C13α-O-deacetylases redelineates the biosynthetic pathway of paclitaxel. bioRxiv 2026. DOI:10.64898/2026.04.28.721278

[28] Chau M, Walker K, Long R, Croteau R. Regioselectivity of taxoid-O-acetyltransferases: Heterologous expression and characterization of a new taxadien-5alpha-ol-O-acetyltransferase. Arch. Biochem. Biophys. 2004, 430, 237-246. DOI:10.1016/j.abb.2004.07.013

[29] Jennewein S, Rithner CD, Williams RM, Croteau R. Taxoid metabolism: Taxoid 14beta-hydroxylase is a cytochrome P450-dependent monooxygenase. Arch. Biochem. Biophys. 2003, 413, 262-270. DOI:10.1016/S0003-9861(03)00090-0

[30] Walker KD, Klettke K, Akiyama T, Croteau R. Cloning, heterologous expression, and characterization of a phenylalanine aminomutase involved in Taxol biosynthesis. J. Biol. Chem. 2004, 279, 53947-53954. DOI:10.1074/jbc.M411215200

[31] Ramirez-Estrada K, Altabella T, Onrubia M, Moyano E, Notredame C, Osuna L, et al. Transcript profiling of jasmonateelicited Taxus cells reveals a beta-phenylalanine-CoA ligase. Plant Biotechnol. J. 2016, 14, 85-96. DOI:10.1111/pbi.12359

[32] Walker K, Long R, Croteau R. The final acylation step in taxol biosynthesis: Cloning of the taxoid C13-side-chain Nbenzoyltransferase from Taxus. Proc. Natl. Acad. Sci. USA 2002, 99, 9166-9171. DOI:10.1073/pnas.082115799

[33] Hefner J, Ketchum REB, Croteau R. Cloning and functional expression of a cDNA encoding geranylgeranyl diphosphate synthase from Taxus canadensis and assessment of the role of this prenyltransferase in cells induced for taxol production. Arch. Biochem. Biophys. 1998, 360, 62-74. DOI:10.1006/abbi.1998.0926

[34] Wildung MR, Croteau R. A cDNA clone for taxadiene synthase, the diterpene cyclase that catalyzes the committed step of taxol biosynthesis. J. Biol. Chem. 1996, 271, 9201-9204. DOI:10.1074/jbc.271.16.9201

[35] Jennewein S, Long RM, Williams RM, Croteau R. Cytochrome P 450 taxadiene 5 alpha-hydroxylase, a mechanistically unusual monooxygenase catalyzing the first oxygenation step of taxol biosynthesis. Chem. Biol. 2004, 11, 379-387. DOI:10.1016/j.chembiol.2004.02.022

[36] Jennewein S, Rithner CD, Williams RM, Croteau RB. Taxol biosynthesis: Taxane 13 alpha-hydroxylase is a cytochrome P450-dependent monooxygenase. Proc. Natl. Acad. Sci. USA 2001, 98, 13595-13600. DOI:10.1073/pnas.251539398

[37] Walker K, Schoendorf A, Croteau R. Molecular cloning of a taxa-4( 20), 11(12)-dien-5alpha-ol-O-acetyl transferase cDNA from Taxus and functional expression in Escherichia coli. Arch. Biochem. Biophys. 2000, 374, 371-380. DOI:10.1006/abbi.1999.1609

[38] Chau M, Jennewein S, Walker K, Croteau R. Taxol biosynthesis: Molecular cloning and of a cytochrome p 450 characterization taxoid 7 beta-hydroxylase. Chem. Biol. 2004, 11, 663-672. DOI:10.1016/S1074-5521(04)00111-5

[39] Chau M, Croteau R. Molecular cloning and characterization of a cytochrome P 450 taxoid 2 alpha-hydroxylase involved in Taxol biosynthesis. Arch. Biochem. Biophys. 2004, 427, 48-57. DOI:10.1016/j.abb.2004.04.016

[40] Walker K, Croteau R. Taxol biosynthesis: Molecular cloning of a benzoyl-CoA. Taxane 2 alpha-O-benzoyltransferase cDNA from Taxus and functional expression in Escherichia coli. Proc. Natl. Acad. Sci. USA 2000, 97, 13591-13596. DOI:10.1073/pnas.250491997

[41] Walker K, Croteau R. Molecular cloning of a 10-deacetylbaccatin III-10-O-acetyl transferase cDNA from Taxus and functional expression in Escherichia coli. Proc. Natl. Acad. Sci. USA 2000, 97, 583-587. DOI:10.1073/pnas.97.2.583

[42] Sanchez-Munoz R, Perez-Mata E, Almagro L, Cusido RM, Bonfill M, Palazon J, et al. A Novel Hydroxylation Step in the Taxane Biosynthetic Pathway: A New Approach to Paclitaxel Production by Synthetic Biology. Front. Bioeng. Biotechnol. 2020, 8, 410. DOI:10.3389/fbioe.2020.00410

[43] Li T, Li B, Liao C, Zhang H, Wang L, Fu T, et al. Transcriptome analysis provides insights into light condition effect on paclitaxel biosynthesis in yew saplings. BMC Plant Biol. 2022, 22, 577. DOI:10.1186/s12870-022-03958-2

[44] Mubeen S, Li ZL, Huang QM, He CT, Yang ZY. Comparative Transcriptome Analysis Revealed the Tissue-Specific Accumulations of Taxanes among Three Experimental Lines of Taxus yunnanensis. J. Agric. Food Chem. 2018, 66, 10410-10420. DOI:10.1021/acs.jafc.8b03502

[45] Wang T, Chen Y, Zhuang W, Zhang F, Shu X, Wang Z, et al. Transcriptome Sequencing Reveals Regulatory Mechanisms of Taxol Synthesis in Taxus wallichiana var. Mairei. Int. J. Genom. 2019, 2019, 1596895. DOI:10.1155/2019/1596895

[46] Yu C, Luo X, Zhang C, Xu X, Huang J, Chen Y, et al. Tissue-specific study across the stem of Taxus media identifies a phloem-specific TmMYB3 involved in the transcriptional regulation of paclitaxel biosynthesis. Plant J. 2020, 103, 95-110. DOI:10.1111/tpj.14710

[47] Yu C, Hou K, Zhang H, Liang X, Chen C, Wang Z, et al. Integrated mass spectrometry imaging and single-cell transcriptome atlas strategies provide novel insights into taxoid biosynthesis and transport in Taxus mairei stems. Plant J. 2023, 115, 1243-1260. DOI:10.1111/tpj.16315

[48] Huang Q, Roessner CA, Croteau R, Scott AI. Engineering Escherichia coli for the Synthesis of Taxadiene, a Key Intermediate in the Biosynthesis of Taxol. Bioorganic Med. Chem. 2001, 9, 2237-2242. DOI:10.1016/S0968-0896(01)00072-4

[49] Zhou K, Qiao K, Edgar S, Stephanopoulos G. Distributing a metabolic pathway among a microbial consortium enhances production of natural products. Nat. Biotechnol. 2015, 33, 377-383. DOI:10.1038/nbt.3095

[50] Boghigian BA, Armando J, Salas D, Pfeifer BA. Computational identification of gene over-expression targets for metabolic engineering of taxadiene production. Appl. Microbiol. Biotechnol. 2012, 93, 2063-2073. DOI:10.1007/s00253-011-3725-1

[51] Abdallah II, Pramastya H, van Merkerk R, Sukrasno, Quax WJ. Metabolic Engineering of Bacillus subtilis Toward Taxadiene Biosynthesis as the First Committed Step for Taxol Production. Front. Microbiol. 2019, 10, 218. DOI:10.3389/fmicb.2019.00218

[52] Biggs BW, Lim CG, Sagliani K, Shankar S, Stephanopoulos G, De Mey M, et al.Overcoming heterologous protein interdependency to optimize P450-mediated Taxol precursor synthesis in Escherichia coli. Proc. Natl. Acad. Sci. USA 2016, 113, 3209-3214. DOI:10.1073/pnas.1515826113

[53] Mutanda I, Li J, Xu F, Wang Y. Recent Advances in Metabolic Engineering, Protein Engineering, and Transcriptome-Guided Insights Toward Synthetic Production of Taxol. Front. Bioeng. Biotechnol. 2021, 9, 632269. DOI:10.3389/fbioe.2021.632269

[54] Ding MZ, Yan HF, Li LF, Zhai F, Shang LQ, Yin Z, et al. Biosynthesis of Taxadiene in Saccharomyces cerevisiae: Selection of geranylgeranyl diphosphate synthase directed by a computer-aided docking strategy. PLoS ONE 2014, 9, e109348. DOI:10.1371/journal.pone.0109348

[55] Nowrouzi B, Li RA, Walls LE, d’Espaux L, Malci K, Liang L, et al. Enhanced production of taxadiene in Saccharomyces cerevisiae. Microb. Cell Factories 2020, 19, 200. DOI:10.1186/s12934-020-01458-2

[56] Walls LE, Malci K, Nowrouzi B, Li RA, d’Espaux L, Wong J, et al. Optimizing the biosynthesis of oxygenated and acetylated Taxol precursors in Saccharomyces cerevisiae using advanced bioprocessing strategies. Biotechnol. Bioeng. 2021, 118, 279-293. DOI:10.1002/bit.27569

[57] Nowrouzi B, Lungang L, Rios-Solis L. Exploring optimal Taxol(R) CYP725A4 activity in Saccharomyces cerevisiae. Microb. Cell Factories 2022, 21, 197. DOI:10.1186/s12934-022-01922-1

[58] Jiang Z, Kempinski C, Bush CJ, Nybo SE, Chappell J. Engineering Triterpene and Methylated Triterpene Production in Plants Provides Biochemical and Physiological Insights into Terpene Metabolism. Plant Physiol. 2016, 170, 702-716. DOI:10.1104/pp.15.01548

[59] Besumbes O, Sauret-Gueto S, Phillips MA, Imperial S, Rodriguez-Concepcion M, Boronat A. Metabolic engineering of isoprenoid biosynthesis in Arabidopsis for the production of taxadiene, the first committed precursor of Taxol. Biotechnol. Bioeng. 2004, 88, 168-175. DOI:10.1002/bit.20237

[60] Rontein D, Onillon S, Herbette G, Lesot A, Werck-Reichhart D, Sallaud C, et al. CYP725A4 from yew catalyzes complex structural rearrangement of taxa-4(5),11(12)-diene into the cyclic ether 5(12)-oxa-3(11)-cyclotaxane. J. Biol. Chem. 2008, 283, 6067-6075. DOI:10.1074/jbc.M708950200

[61] Li J, Mutanda I, Wang K, Yang L, Wang J, Wang Y. Chloroplastic metabolic engineering coupled with isoprenoid pool enhancement for committed taxanes biosynthesis in Nicotiana benthamiana. Nat. Commun. 2019, 10, 4850. DOI:10.1038/s41467-019-12879-y

[62] Fu J, Xu W, Huang W, Wang B, Li S, Zhang J, et al. Importation of taxadiene synthase into chloroplast improves taxadiene production in tobacco. Planta 2021, 253, 107. DOI:10.1007/s00425-021-03626-z

[63] He J, Plácido JPA, Pateraki I, Kampranis S, Favero BT, Lütken H. Hairy root induction of Taxus baccata L. by natural transformation with Rhizobium rhizogenes. Horticulturae 2023, 9, 4. DOI:10.3390/horticulturae9010004

[64] Ajikumar PK, Xiao WH, Tyo KE, Wang Y, Simeon F, Leonard E, et al. Isoprenoid pathway optimization for Taxol precursor overproduction in Escherichia coli. Science 2010, 330, 70-74. DOI:10.1126/science.1191652

[65] Boghigian BA, Salas D, Ajikumar PK, Stephanopoulos G, Pfeifer BA. Analysis of heterologous taxadiene production in K- and B-derived Escherichia coli. Appl. Microbiol. Biotechnol. 2012, 93, 1651-1661. DOI:10.1007/s00253-011-3528-4

[66] Wu Q-Y, Huang Z-Y, Wang J-Y, Yu H-L, Xu J-H. Construction of an Escherichia coli cell factory to synthesize taxadien-5α-ol, the key precursor of anti-cancer drug paclitaxel. Bioresour. Bioprocess. 2022, 9, 82. DOI:10.1186/s40643-022-00569-5

[67] Wang J-Y, Huang Z-Y, Wu Q-Y, Pan J, Li C-X, Xu J-H. Facile biosynthesis of taxadiene by a newly constructed Escherichia coli strain fusing enzymes taxadiene synthase and geranylgeranyl pyrophosphate synthase. Process Biochem. 2022, 122, 129-136. DOI:10.1016/j.procbio.2022.09.005

[68] Engels B, Dahm P, Jennewein S. Metabolic engineering of taxadiene biosynthesis in yeast as a first step towards Taxol (Paclitaxel) production. Metab. Eng. 2008, 10, 201-206. DOI:10.1016/j.ymben.2008.03.001

[69] Walls LE, Martinez JL, Rios-Solis L. Enhancing Saccharomyces cerevisiae Taxane Biosynthesis and Overcoming Nutritional Stress-Induced Pseudohyphal Growth. Microorganisms 2022, 10, 163. DOI:10.3390/microorganisms10010163

[70] Xu M, Xie W, Luo Z, Li CX, Hua Q, Xu J. Improving solubility and copy number of taxadiene synthase to enhance the titer of taxadiene in Yarrowia lipolytica. Synth. Syst. Biotechnol. 2023, 8, 331-338. DOI:10.1016/j.synbio.2023.04.002

[71] Min L, Han JC, Zhang W, Gu CC, Zou YP, Li CC. Strategies and Lessons Learned from Total Synthesis of Taxol. Chem. Rev. 2023, 123, 4934-4971. DOI:10.1021/acs.chemrev.2c00763

[72] Hasan MM, Kim HS, Jeon JH, Kim SH, Moon B, Song JY, et al. Metabolic engineering of Nicotiana benthamiana for the increased production of taxadiene. Plant Cell Rep. 2014, 33, 895-904. DOI:10.1007/s00299-014-1568-9

[73] Wick C, Somani A, Liu JC, Karunadasa SS, Xu SL, Fordyce PM, et al. FoTO1 orchestrates Taxol biosynthesis through catalytic and non-catalytic mechanisms. BioRxiv 2026. DOI:10.64898/2026.03.21.713420

[74] Wang Y, Guo B, Miao Z, Tang K. Transformation of taxol-producing endophytic fungi by restriction enzyme-mediated integration (REMI). FEMS Microbiol. Lett. 2007, 273, 253-259. DOI:10.1111/j.1574-6968.2007.00801.x

[75] El-Sayed ASA, Abdel-Ghany SE, Ali GS. Genome editing approaches: Manipulating of lovastatin and taxol synthesis of filamentous fungi by CRISPR/Cas 9 system. Appl. Microbiol. Biotechnol. 2017, 101, 3953-3976. DOI:10.1007/s00253-017-8263-z

[76] Zhong J, Wang Y, Chen Z, Yalikun Y, He L, Liu T, et al. Engineering cyanobacteria as a new platform for producing taxol precursors directly from carbon dioxide. Biotechnol. Biofuels Bioprod. 2024, 17, 99. DOI:10.1186/s13068-024-02555-9

[77] Ansbacher T, Freud Y, Major DT. Slow-Starter Enzymes: Role of Active-Site Architecture in the Catalytic Control of the Biosynthesis of Taxadiene by Taxadiene Synthase. Biochemistry 2018, 57, 3773-3779. DOI:10.1021/acs.biochem.8b00452

[78] Liu H, Chen Q. Computational protein design for given backbone: Recent progresses in general method-related aspects. Curr. Opin. Struct. Biol. 2016, 39, 89-95. DOI:10.1016/j.sbi.2016.06.013

[79] Koksal M, Jin Y, Coates RM, Croteau R, Christianson DW. Taxadiene synthase structure and evolution of modular architecture in terpene biosynthesis. Nature 2011, 469, 116-120. DOI:10.1038/nature09628

[80] Soliman S, Tang Y. Natural and engineered production of taxadiene with taxadiene synthase. Biotechnol. Bioeng. 2015, 112, 229-235. DOI:10.1002/bit.25468

[81] Van Rijn JPM, Escorcia AM, Thiel W. QM/MM study of the taxadiene synthase mechanism. J. Comput. Chem. 2019, 40, 1902-1910. DOI:10.1002/jcc.25846

[82] Kanda Y, Ishihara Y, Wilde NC, Baran PS. Two-Phase Total Synthesis of Taxanes: Tactics and Strategies. J. Org. Chem. 2020, 85, 10293-10320. DOI:10.1021/acs.joc.0c01287

[83] Barton NA, Marsh BJ, Lewis W, Narraidoo N, Seymour GB, Fray R, et al. Accessing low-oxidation state taxanes: Is taxadiene-4(5)-epoxide on the taxol biosynthetic pathway? Chem. Sci. 2016, 7, 3102-3107. DOI:10.1039/C5SC03463A

[84] Schrepfer P, Buettner A, Goerner C, Hertel M, van Rijn J, Wallrapp F, et al. Identification of amino acid networks governing catalysis in the closed complex of class I terpene synthases. Proc. Natl. Acad. Sci. USA 2016, 113, 958-967. DOI:10.1073/pnas.1519680113

[85] Yadav VG. Unraveling the multispecificity and catalytic promiscuity of taxadiene monooxygenase. J. Mol. Catal. B Enzym. 2014, 110, 154-164. DOI:10.1016/j.molcatb.2014.10.004

[86] Song X, Wang Q, Zhu X, Fang W, Liu X, Shi C, et al. Unraveling the Catalytic Mechanism of Taxadiene-5α-hydroxylase from Crystallography and Computational Analyses. ACS Catal. 2024, 14, 3912-3925. DOI:10.1021/acscatal.3c05807

[87] Zhang MF, Xie WL, Chen C, Li CX, Xu JH. Computational redesign of taxane-10β-hydroxylase for de novo biosynthesis of a key paclitaxel intermediate. Appl. Microbiol. Biotechnol. 2023, 107, 7105-7117. DOI:10.1007/s00253-023-12784-x

[88] Li BJ, Wang H, Gong T, Chen JJ, Chen TJ, Yang JL, et al. Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production. Nat. Commun. 2017, 8, 15544. DOI:10.1038/ncomms15544

[89] Lin SL, Wei T, Lin JF, Guo LQ, Wu GP, Wei JB, et al. Bio-production of Baccatin III, an Important Precursor of Paclitaxel by a Cost-Effective Approach. Mol. Biotechnol. 2018, 60, 492-505. DOI:10.1007/s12033-018-0090-7

[90] Chen JJ, Liang X, Wang F, Wen YH, Chen TJ, Liu WC, et al. Combinatorial mutation on the β-glycosidase specific to 7-β-xylosyltaxanes and increasing the mutated enzyme production by engineering the recombinant yeast. Acta Pharm. Sin. B 2019, 9, 626-638. DOI:10.1016/j.apsb.2018.11.003

[91] Bai J, Li J, Zhang Y, Chang H, Zhang N, Liu Y, et al. FoTO1 is an epoxide isomerase in paclitaxel biosynthesis. BioRxiv 2026. DOI:10.64898/2026.03.30.715439

[92] Zhu X, Liu X, Liu T, Wang Y, Ahmed N, Li Z, et al. Synthetic biology of plant natural products: From pathway elucidation to engineered biosynthesis in plant cells. Plant Commun. 2021, 2, 100229. DOI:10.1016/j.xplc.2021.100229

[93] Li J, Liu X, Zhu X, Liu J, Zhang L, Ahmed N, et al. Biochemical synthesis of taxanes from mevalonate. Synth. Syst. Biotechnol. 2024, 9, 694-700. DOI:10.1016/j.synbio.2024.05.002