| Targets |
The molecular target of 4-Methylumbelliferyl beta-D-cellobioside is the enzyme (1,4)-beta-glucanase, also known as cellulase. More specifically, it is a substrate for endo- and exo-(1,4)-beta-glucanases. These enzymes catalyze the cleavage of the beta-1,4-glycosidic bonds in cellulose. The substrate is not a drug, so it does not inhibit or activate the enzyme; instead, it is a probe that is cleaved by the enzyme. The rate of hydrolysis is a direct measure of the enzyme's catalytic activity.
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| ln Vitro |
In vitro, 4-Methylumbelliferyl beta-D-cellobioside (MUC) is used as a substrate to evaluate the reducing-end exo-activity of diverse cellulases. Its "activity" is its susceptibility to hydrolysis by cellulases. The rate of hydrolysis is measured by the increase in fluorescence. It has no EC₅0 or IC₅0 for a therapeutic effect. It is used to determine the kinetic parameters (Km, Vmax) of cellulase enzymes. For example, it was used to assay cellulase from Acetivibrio cellulolyticus. It is also used for activity staining of cellulases in polyacrylamide gels.
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| ln Vivo |
No in vivo activity has been reported for 4-Methylumbelliferyl beta-D-cellobioside as it is not a drug. It is a research tool that is used to measure enzyme activity in vitro or in situ. In live organisms, it could theoretically be used as an imaging probe if the target enzyme is present, but such applications are not standard. Its primary use is in biochemical and biomass conversion research, not in animal models of disease.
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| Enzyme Assay |
No enzyme/receptor binding assays are performed; instead, the compound is used in enzyme kinetics. In a typical assay, a buffer solution (e.g., 50 mM sodium citrate, pH 5.0) containing 50-200 uM of the substrate is prepared. An aliquot of a cellulase enzyme sample (e.g., from Clostridium thermocellum or Acetivibrio cellulolyticus) is added to the substrate solution in a 96-well plate. The reaction mixture is incubated at a specific temperature (e.g., 50degC) for a defined time (e.g., 5-30 minutes). The reaction is stopped by adding a basic stop solution (e.g., 1 M Na2CO3) to raise the pH, which also maximizes the fluorescence of the released 4-MU product. Fluorescence is measured (excitation ~360 nm, emission ~450 nm). A standard curve of 4-MU is used to quantify the amount of product formed.
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| Cell Assay |
No cell-based biological assays for viability or proliferation are performed with this substrate. Its use in cell culture would be to measure the activity of cellulases, which are not typically produced by mammalian cells. In a typical protocol, a microbial culture producing cellulases is grown. The culture supernatant containing the secreted enzymes is collected. An aliquot of the supernatant is then mixed with the substrate solution in a buffer (e.g., 50 mM sodium citrate, pH 5.0). After incubation, the reaction is stopped, and fluorescence is measured as described in the cell-free protocol. This provides a rapid and sensitive measure of the enzyme activity in the microbial culture.
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| Animal Protocol |
No animal experiments are performed with 4-Methylumbelliferyl beta-D-cellobioside as it is not a drug. It is a biochemical reagent used for in vitro enzyme assays. It is not administered to animals. In research, it can be used to analyze enzyme activity from samples collected from animals, such as analyzing the gut microbiome of herbivores. The gut contents are collected post-mortem, and the extract is used in the cell-free enzyme assay described previously to measure the presence and activity of cellulase-producing bacteria.
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| ADME/Pharmacokinetics |
No pharmacokinetic data for 4-Methylumbelliferyl beta-D-cellobioside are available, as it is not a drug. Its physical properties are known: it is a white solid, soluble in water and organic solvents like DMSO. It is stable when stored at -20degC. The compound is a fluorogenic substrate, meaning its fluorescence is negligible until it is hydrolyzed. Upon enzymatic cleavage, it releases 4-methylumbelliferone, which has a large fluorescence quantum yield and a visible blue fluorescence. It is not intended for systemic administration.
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| Toxicity/Toxicokinetics |
The toxicity of 4-Methylumbelliferyl beta-D-cellobioside has not been evaluated for therapeutic purposes. It is considered a low-hazard laboratory chemical. Standard laboratory safety precautions should be taken: wear gloves, a lab coat, and eye protection. Avoid breathing dust. The hydrolysis product, 4-methylumbelliferone (4-MU), has been evaluated and is considered to have low toxicity. The compound is not a drug, and there are no human safety data. It is strictly for research use only, not for diagnostic or therapeutic use.
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| References | |
| Additional Infomation |
4-Methylumbelliferone β-D-cellobiose glycoside belongs to the coumarin class of glycosides. It is a substrate of cellobiose hydrolase I.
4-Methylumbelliferyl beta-D-cellobioside is not a drug and has no clinical trials or regulatory approval. It is a research-grade biochemical used as a fluorogenic substrate for cellulase assays. It is particularly useful for measuring the activity of reducing-end exo-glucanases. The assay is highly sensitive, allowing for kinetic studies of cellulolytic enzymes in vitro. It is also used for activity staining of cellulases in polyacrylamide gels, making it an essential tool in biomass conversion research and the study of microbial degradation of cellulose. It is stored at -20degC. |
| Exact Mass |
500.153
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|---|---|
| CAS # |
72626-61-0
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| PubChem CID |
126287
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| Appearance |
White to off-white solid powder
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| Density |
1.65 g/cm3
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| Boiling Point |
830.3ºC at 760 mmHg
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| Melting Point |
230ºC
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| Flash Point |
286ºC
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| Index of Refraction |
1.677
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| LogP |
0
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| Hydrogen Bond Donor Count |
7
|
| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
35
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| Complexity |
776
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| Defined Atom Stereocenter Count |
10
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| InChi Key |
PRTGXBPFDYMIJH-MKQZUAMYSA-N
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| InChi Code |
InChI=1S/C22H28O13/c1-8-4-14(25)32-11-5-9(2-3-10(8)11)31-21-19(30)17(28)20(13(7-24)34-21)35-22-18(29)16(27)15(26)12(6-23)33-22/h2-5,12-13,15-24,26-30H,6-7H2,1H3/t12-,13-,15-,16+,17-,18-,19-,20-,21-,22+/m1/s1
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| Chemical Name |
7-[(2S,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-4-methylchromen-2-one
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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 |
| 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) |
DMSO : ≥ 125 mg/mL (~249.78 mM)
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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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