Integrating Pathway Engineering and FoTO1-Mediated Oxidation for Taxadien-5α-ol Production
- 1State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy
- 2China Pharmaceutical University
- 3Public Laboratory Platform
Abstract
Microbial reconstruction of paclitaxel biosynthesis is hindered by limited taxadiene (1) supply and poor selectivity in the first oxidative step. Here, we engineered Escherichia coli for the efficient de novo production of taxadien-5α-ol (2) by integrating enhanced taxadiene (1) biosynthesis with selective oxidation. Coordinated engineering of taxadiene synthase expression, precursor supply, pathway regulation, and geranylgeranyl diphosphate synthase increased the titer of 1 7.5-fold, from 20.3 to 152.6 mg L−1. However, the CYP725A4–cytochrome P450 reductase fusion produced only 2.2 mg L−1 of 2, representing 7.4% of the detected oxygenated taxanes. Coexpression of facilitator of taxane oxidation 1 (FoTO1) markedly redirected the oxidation profile toward 2, increasing its proportion to 94.2% and its titer 20.8-fold to 45.5 mg L−1. Subsequent fermentation optimization increased the titer to 74.0 mg L−1, and addition of a 15% (v/v) n-dodecane overlay further increased production to 179.9 mg L−1. To our knowledge, this represents the highest reported titer of individually quantified taxadien-5α-ol (2) in a heterologous microbial host. These results highlight the importance of coupling precursor supply with control of oxidation selectivity and establish an efficient platform for further reconstruction of microbial paclitaxel biosynthesis.
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BibTeX
@misc{li2026integrating,
title = {{Integrating Pathway Engineering and FoTO1-Mediated Oxidation for Taxadien-5α-ol Production}},
author = {Haixin Li and Shuyao Xu and Mingyu Yue and Lianchun Deng and Hui-Min Xu and Hongwei Chen and Liao-Bin Dong},
howpublished = {ChemRxiv},
year = {2026},
month = sep,
doi = {10.26434/chemrxiv.15009335/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009335/v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Integrating Pathway Engineering and FoTO1-Mediated Oxidation for Taxadien-5α-ol Production
AU - Haixin Li
AU - Shuyao Xu
AU - Mingyu Yue
AU - Lianchun Deng
AU - Hui-Min Xu
AU - Hongwei Chen
AU - Liao-Bin Dong
PY - 2026
DA - 2026/09/23
PB - ChemRxiv
DO - 10.26434/chemrxiv.15009335/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009335/v1
AB - Microbial reconstruction of paclitaxel biosynthesis is hindered by limited taxadiene (1) supply and poor selectivity in the first oxidative step. Here, we engineered Escherichia coli for the efficient de novo production of taxadien-5α-ol (2) by integrating enhanced taxadiene (1) biosynthesis with selective oxidation. Coordinated engineering of taxadiene synthase expression, precursor supply, pathway regulation, and geranylgeranyl diphosphate synthase increased the titer of 1 7.5-fold, from 20.3 to 152.6 mg L−1. However, the CYP725A4–cytochrome P450 reductase fusion produced only 2.2 mg L−1 of 2, representing 7.4% of the detected oxygenated taxanes. Coexpression of facilitator of taxane oxidation 1 (FoTO1) markedly redirected the oxidation profile toward 2, increasing its proportion to 94.2% and its titer 20.8-fold to 45.5 mg L−1. Subsequent fermentation optimization increased the titer to 74.0 mg L−1, and addition of a 15% (v/v) n-dodecane overlay further increased production to 179.9 mg L−1. To our knowledge, this represents the highest reported titer of individually quantified taxadien-5α-ol (2) in a heterologous microbial host. These results highlight the importance of coupling precursor supply with control of oxidation selectivity and establish an efficient platform for further reconstruction of microbial paclitaxel biosynthesis.
ER -