Taxadiene Analysis Service - Creative Proteomics

Service Details Q&A DemoCase Study

What are taxadiene?

Taxadiene (C20H32), a key diterpene hydrocarbon, serves as the pivotal biosynthetic precursor for Taxol. Its biosynthesis begins with the cyclization of geranylgeranyl pyrophosphate (GGPP) catalyzed by taxadiene synthase (TXS), forming the unique tricyclic scaffold. As the first committed intermediate in Taxol biosynthesis, taxadiene quantification and structural analysis are critical for:

Drug development: Monitoring flux through the Taxol biosynthetic pathway in Taxus species or engineered microbial systems.
Metabolic engineering: Optimizing heterologous production of taxadiene in E. coli or yeast chassis.
Quality control: Ensuring purity in commercial taxadiene batches for pharmaceutical intermediates.

Creative Proteomics delivers taxadiene analysis services tailored to accelerate research and industrial workflows.

Taxadiene Analysis Service by Creative Proteomics

Creative Proteomics provides a wide range of specialized services for Taxadiene analysis to ensure high-precision data back your research and development processes. Our service offerings include:

1. Taxadiene ldentification and Quantification

Targeted GC-MS or LC-MS/Ms analysis for accurate quantification of taxadiene
Absolute and relative quantification with internal standards
Detection sensitivity at trace levels in plant extracts, microbial cultures, or engineered systems

2.Pathway Profiling and Metabolite Analysis

comprehensive metabolic profiling to identify taxadiene biosynthetic intermediates
Monitoring of taxadiene pathway fluxes using stable isotope labeling (SIL)
Comparative analysis of engineered strains for metabolic optimization

3. Structural Characterization

High-resolution MS and MS/MS analysis for taxadiene structural elucidation
Fragmentation pattern analysis to determine molecular structure and confirm identity

4. Isotope Tracing Studies

Quantifying metabolic rates by labelling paclitaxel and other paclitaxel metabolites reveals dynamic metabolic processes.
Combined with mass spectrometry to track the binding of isotopically labelled compounds in metabolic pathways to determine metabolic rates and pathway activity.

5. Custom Assay Development

Development and validation of customized taxadiene quantification assays
Optimization of extraction and sample preparation protocols for diverse sample matrices

6. Data Interpretation and Report Generation

Detailed, scientifically rigorous reports, including all relevant data, trends and analyses
Assist in decision-making and further research

List of Taxadiene-Related Metabolites (including but not limited to)

Detected Substance	Type	Metabolic Pathway
Taxadiene	Diterpene	Biosynthesis of paclitaxel from taxadiene in Taxus species
Taxol (Paclitaxel)	Terpenoid	Conversion from taxadiene via several enzymatic steps
10-deacetylbaccatin III	Precursor to Paclitaxel	Intermediate in the paclitaxel biosynthetic pathway
Baccatin III	Diterpene	Intermediate leading to paclitaxel formation
7-epi-taxol	Taxoid derivative	Side-product in the biosynthesis pathway
CYP450 Enzymes	Enzyme family	Key enzymes involved in hydroxylation steps in taxadiene metabolism
geranylgeranyl pyrophosphate（GGPP）	Precursor	Terpenoid synthesis precursors

Techniques and Instrumentation for Taxadiene Analysis

Bruker 400 MHz NMR spectrometer (Figure from Bruker)

7890B Gas Chromatograph (Figure from Agilent)

ISQ™ EC Single Quadrupole MS (Figure from Thermo Scientific)

Mass-Based 1290 Infinity II Preparative LC/MSD System (Figure from Agilent)

Workflow for Taxadiene Analysis Service

Why Choose Us?

High Precision: We employ state-of-the-art analytical instrumentation, such as LC-MS/MS and GC-MS, to deliver highly accurate quantifications of taxadiene and its metabolites with excellent sensitivity and specificity.
Comprehensive Metabolic Profiling: Our service provides not just a measure of taxadiene but also detailed insights into the entire metabolic pathway leading to its derivatives and final products.
Taxane-Specific Targeted Analytical Platform: Utilizing LC-MS/MS in Multiple Reaction Monitoring mode, this system enables simultaneous quantification of 12 paclitaxel precursor compounds, with a detection sensitivity of 0.1 ng/mL and quantitative accuracy exceeding 98%.
Customization: Tailored services that meet your specific needs, whether you're studying plant-based systems, microbial biosynthesis, or engineered strains for taxadiene production.
Speed and Efficiency: Our advanced systems allow for fast processing times, enabling rapid turnaround without compromising quality.
Scientific Expertise: With over 20 years of experience in biochemical and molecular analysis, our team of experts ensures that your samples are handled with the highest level of scientific integrity.
Perfect after-sales service: Comprehensive interpretation of the final report, free technical advice, and data review service.
Comprehensive analyses: 12 analyses, 22 graphical displays.

Applications of Taxadiene Analysis

Biotechnology and Synthetic Biology

Investigating microbial or plant-based systems engineered for the biosynthesis of taxadiene and its derivatives.

Agricultural Science

Understanding the genetic and environmental factors that influence taxadiene production in Taxus species or other plants

Pharmacology and Drug Development

Studying the pathways that lead to paclitaxel production and optimizing taxadiene biosynthesis for pharmaceutical applications.

Academic Research

Map taxadiene's role in plant defense mechanisms

Sample Requirements for Taxadiene Analysis Assay

Sample Types	Minimum Quantity	Required Conditions
Plant Tissues	500 mg	Freeze-dried or fresh samples, stored at -80°C
Microbial cultures	10-50 mL	Liquid cultures grown under controlled conditions
Plant extracts	2-5 mL	Extracts should be in organic solvents like methanol or ethanol
Cell cultures (suspension)	1-5 mL	Harvest cells in mid-log phase, flash-frozen

Q: How long does a standard taxadiene quantification take?: A: 24–48 hours for GC-MS quantification; 72 hours for full pathway flux analysis.
Q: Can I submit lyophilized samples?: A: Yes, but reconstitute in 500 µL hexane before shipping at -20℃.
Q: What's the LOD for taxadiene in yeast cultures?: A: 0.5 ng/mL (matrix-matched calibration applied).
Q: Do you recommend technical replicates?: A: Yes—we include triplicate runs at no extra cost.
Q: Can I receive raw data files?: A: Yes, .RAW (Thermo), .D (Agilent), and .Wiff (Sciex) formats provided.
Q: Do you analyze taxadiene epoxides?: A: Yes, via LC-QTOF (requires ≥50 µL sample).
Q: How is data confidentiality handled?: A: All data is encrypted, and projects are assigned anonymized IDs.

Chromatogram showing TIC peaks for mono-oxidized and poly-functionalized compounds with GGOH retention time analysis (Figure from Jack Chun-Ting Liu et al., 2024)

Chromatogram displaying intensity vs. time for 10-DAB, BAC, DAP, and PTX compounds with cps values ranging from 1.8E5 to 2.0E4 (Figure from Chunna Yu et al., 2020)

Case. Characterization and heterologous reconstitution of Taxus biosynthetic enzymes leading to baccatin III

Background:

Gene expression analysis of candidate genes and paclitaxel biosynthetic genes.
Metabolite extraction and LC-MS analysis.
This analysis aims to identify two key deletion genes in the paclitaxel synthesis pathway, TOT and T9αH, to reveal a novel mechanism by which plant cells catalyze the formation of oxetane structures.

Samples:

Different tissues of Picea abies (needles, bark and roots of females (F) and males (M)), Nicotiana benthamiana, freeze-dried tissues.
Three biological replicates were included for each line, with six plants for each biological replicate

Technical methods procedure:

Expression levels of candidate genes and paclitaxel biosynthesis genes were analyzed based on RNA-seq data.
Taxus mairei, Nicotiana benthamiana and insect cells were freeze-dried.
Metabolites were extracted by sonication of freeze-dried tissues with 1 mL MeOH for 20 min at room temperature.
Centrifugation to obtain supernatant
Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) was used for qualitative analysis of metabolites.
Metabolites were quantified using standard curves.
Concentration determination of paclitaxel metabolite content using an ion chromatography system.

Results

Screening and validation of oxidase synthesis genes in taxadiene metabolism.
Determination of changes in the content of intermediates in the paclitaxel synthesis pathway.
Identification of a bifunctional cytochrome P450 enzyme [paclitaxel oxetane enzyme 1 (TOT1)] in Taxus mairei that catalyzes an oxidative rearrangement in paclitaxel oxetane formation.
The biosynthetic pathway of baccatin III was successfully artificially reconstructed in tobacco by co-expressing other known genes involved in baccatin III synthesis.

The proposed biosynthetic pathway of paclitaxel. (Figure from Bin Jiang et al., 2024)

References

Bin Jiang et al., Characterization and heterologous reconstitution of Taxus biosynthetic enzymes leading to baccatin III.Science383,622-629(2024). https://doi.org/10.1126/science.adj3484
Li, J., Mutanda, I., Wang, K. et al. Chloroplastic metabolic engineering coupled with isoprenoid pool enhancement for committed taxanes biosynthesis in Nicotiana benthamiana. Nat Commun 10, 4850 (2019). https://doi.org/10.1038/s41467-019-12879-y.
Jin-Yi Wang, Zheng-Yu Huang. et al. Facile biosynthesis of taxadiene by a newly constructed Escherichia coli strain fusing enzymes taxadiene synthase and geranylgeranyl pyrophosphate synthase.Process Biochemistry 122, 2 (2022). https://doi.org/10.1016/j.procbio.2022.09.005

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