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| 1mg |
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| Other Sizes |
| Targets |
Shikimate-3-phosphate lithium targets the active site of EPSP synthase (5-enolpyruvylshikimate-3-phosphate synthase, EC 2.5.1.19). This enzyme catalyzes the transfer of the enolpyruvyl moiety from phosphoenolpyruvate (PEP) to shikimate-3-phosphate (S3P) to form EPSP. The compound serves as a substrate, binding to the enzyme together with PEP. In the presence of glyphosate, glyphosate competes with PEP and forms a ternary complex with EPSP synthase and S3P, inhibiting the enzyme. Thus, S3P is an indispensable tool for studying glyphosate resistance mechanisms and enzyme catalysis.
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| ln Vitro |
In vitro, shikimate-3-phosphate lithium is used to measure EPSP synthase activity. In a standard assay, the reaction mixture contains 50 mM HEPES (pH 7.5), 1 mM S3P, 1 mM PEP, 1 mM ammonium molybdate, and 1 ug of purified EPSP synthase in a final volume of 100 uL. After incubation at 25degC for 5-20 min, the reaction is stopped by adding malachite green reagent, which forms a color complex with released phosphate. The absorbance at 620 nm is measured. The specific activity (umol/min/mg) is calculated. Alternatively, the formation of EPSP can be quantified by HPLC. The Km of S3P for EPSP synthase is typically in the range of 10-50 uM. Glyphosate inhibition is assessed by pre-incubating the enzyme with S3P and varying concentrations of glyphosate.
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| ln Vivo |
Shikimate-3-phosphate lithium is not used as a therapeutic agent in vivo because humans lack the shikimate pathway. However, it can be administered to plants or bacteria to study metabolic flux. For example, in glyphosate-treated plants, exogenous S3P can partially overcome growth inhibition by competing with the herbicide, but this is not a practical use. In animal studies, S3P is not administered due to lack of target. The compound is strictly a research tool for studying plant physiology, bacterial metabolism, and herbicide mechanism. No in vivo pharmacological activity exists.
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| Enzyme Assay |
A non-cellular assay for S3P is the EPSP synthase phosphate release assay described above. Additionally, the purity and identity of the compound can be confirmed by 3¹P NMR spectroscopy (phosphate ester peak around 4-5 ppm), by HPLC on an anion-exchange column (e.g., Dionex CarboPac) with pulsed amperometric detection, or by LC-MS (negative ion mode, m/z for shikimate-3-phosphate [M-H]- = 255.0). The lithium salt is used because it is more soluble and stable than the free acid. A colorimetric test for free phosphate using malachite green can be used to check for contamination. Standard curves are prepared using known concentrations of S3P.
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| Cell Assay |
Shikimate-3-phosphate lithium is not used in mammalian cell assays because mammalian cells do not have the shikimate pathway. However, it can be used in bacterial cell extracts or permeabilized bacterial cells to measure EPSP synthase activity. For instance, E. coli cells are grown, harvested, and lysed by sonication. The crude lysate is incubated with S3P, PEP, and cofactors, and the production of EPSP is measured by HPLC. Alternatively, genetically engineered yeast cells expressing plant EPSP synthase can be used. S3P is also used as a positive control in glyphosate binding assays. In plant cell cultures (e.g., carrot or Arabidopsis), radiolabeled S3P ([14C]) can be added to track carbon flow through the shikimate pathway.
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| Animal Protocol |
In vivo animal experiments are not performed with shikimate-3-phosphate because it is not absorbed or metabolized by animals. However, plant studies are relevant: For example, glyphosate-resistant transgenic plants (e.g., Roundup Ready soybeans) are treated with glyphosate, and the accumulation of S3P in leaf tissues is measured by LC-MS/MS as an indicator of EPSP synthase inhibition. In non-transgenic plants, S3P levels rise dramatically after glyphosate treatment due to the block in the pathway. Exogenous application of S3P (1-10 mM via leaf infiltration) can rescue glyphosate-induced growth inhibition in some plant species, demonstrating that the compound can enter plant cells and serve as a substrate. This is used to study resistance mechanisms.
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| ADME/Pharmacokinetics |
Shikimate-3-phosphate is an anionic, highly polar molecule that does not readily cross cell membranes. In plants, it is localized in the plastid where the shikimate pathway resides. Exogenously applied S3P is poorly absorbed; thus, no typical pharmacokinetic parameters (like Cmax, t1/2) are determined. In bacteria, S3P is taken up via specific phosphate transporters. For research, the compound is typically used at high concentrations (0.1-5 mM) in vitro. The lithium salt improves solubility compared to the sodium or ammonium salt. There are no published ADME studies because S3P is not a drug candidate; it is an intermediate. It is stable in solution at neutral pH but can be hydrolyzed at low or high pH.
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| Toxicity/Toxicokinetics |
Shikimate-3-phosphate is non-toxic to animals because animals do not have the shikimate pathway and cannot metabolize it. The LD50 is likely >2000 mg/kg (oral) in rats. However, lithium salt introduces lithium ions, which at high doses can cause lithium toxicity (nausea, tremors, renal impairment). But in typical assays, the amount of lithium is negligible (e.g., 1 mM S3P contains 1 mM Li+, which is far below therapeutic lithium levels). The compound should be handled as a general laboratory chemical. No specific toxicity studies have been published. Use standard precautions: avoid inhalation and skin contact. The free acid form may be an irritant.
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| References | |
| Additional Infomation |
Shikimate-3-phosphate lithium (no formal CAS number is readily available; the free acid CAS is 29908-08-1, but the lithium salt is often custom synthesized) is a research biochemical. Molecular formula of free acid: C₇H11O₈P (MW 254.13). The lithium salt typically contains 1-2 Li atoms. It is a white powder, soluble in water. The compound is used in mechanistic studies of EPSP synthase and for glyphosate inhibition assays. It is also used in the synthesis of labeled EPSP for metabolic flux analysis. This compound is not a drug and has no clinical applications. It is strictly for research use. Suppliers offer it as part of herbicide resistance kits. Store at -20degC, desiccated, avoid freeze-thaw cycles.
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| Molecular Formula |
C7H11O8P.XLI
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| Molecular Weight |
254.13 (free base)
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| Appearance |
Solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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