| Size | Price | Stock | Qty |
|---|---|---|---|
| 1g |
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| 10g |
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| Other Sizes |
| Targets |
L-gulono-1,4-lactone dehydrogenase (Rv1771, Mycobacterium tuberculosis, \(K_m = 5.5\) mM as substrate)
Cytochrome c (exogenous electron acceptor, \(K_m = 4.7\) μM) [1] |
|---|---|
| ln Vitro |
L-Gulono-1,4-lactone is specifically oxidized by the recombinant Rv1771 protein (L-gulono-1,4-lactone dehydrogenase) from Mycobacterium tuberculosis, with no activity towards L-galactono-1,4-lactone, D-glucurono-3,6-lactone, D-glucuronic acid, D-arabinose, or D-xylose [1]
The oxidation reaction uses cytochrome c or phenazine methosulfate as exogenous electron acceptors (phenazine methosulfate is ~3 times more efficient); molecular oxygen does not serve as an electron acceptor [1] The reaction follows Michaelis–Menten kinetics, with a \(V_{max}\) of \(2.44 \mu mol \cdot h^{-1} \cdot mg protein^{-1}\); optimal pH is 7.5–8, and optimal temperature is 39°C [1] The reaction is completely inhibited by 1 mM N-ethylmaleimide, \(Cu^{2+}\), \(Zn^{2+}\), and potassium cyanide; 1 mM EDTA inactivates the enzyme (requires trace divalent metal ions); \(Mg^{2+}\) and \(Ca^{2+}\) have no effect, while 1 mM \(Mn^{2+}\) causes 21% inhibition [1] L-gulono-1,4-lactone dehydrogenase activity (0.17 mU·mg protein⁻¹) is detected in the soluble fraction of Mycobacterium bovis BCG crude extracts [1] |
| Enzyme Assay |
Dehydrogenase activity assay with cytochrome c as electron acceptor: The reaction mixture (1 mL) contains 25 mM L-Gulono-1,4-lactone, 0.121 mM cytochrome c, and affinity-purified Rv1771 enzyme in 50 mM phosphate buffer (pH 7.3). Dithiothreitol and glutathione are removed by gel filtration before assay. Activity is measured spectrophotometrically at 550 nm by monitoring the L-gulono-1,4-lactone-dependent reduction of cytochrome c [1]
Dehydrogenase activity assay with phenazine methosulfate as electron acceptor: The reaction mixture includes 25 mM L-Gulono-1,4-lactone, 2.5 mM phenazine methosulfate, 100 μM 2,6-dichloroindophenol, and the enzyme in 50 mM phosphate buffer (pH 7.3). Activity is determined by measuring the decrease in absorbance at 610 nm due to the reduction of 2,6-dichloroindophenol [1] |
| Cell Assay |
Crude enzyme extract preparation from M. bovis BCG: M. bovis BCG cultures are grown to mid-exponential phase, collected by centrifugation, resuspended in 100 mM phosphate buffer (pH 7.3) containing 1 mM phenylmethanesulfonyl fluoride, and disrupted by sonication. Unbroken cells are removed by low-speed centrifugation, and the supernatant is further centrifuged at 25,000 g to separate soluble extract and insoluble cell envelope fraction. The soluble fraction is used to measure L-gulono-1,4-lactone dehydrogenase activity with L-Gulono-1,4-lactone as substrate [1]
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| ADME/Pharmacokinetics |
Biosynthetic precursor: L-gulonic acid-1,4-lactone can be generated by C1 reduction of D-glucuronic acid or D-glucuronic acid-3,6-lactone; NADPH-dependent D-glucuronic acid-3,6-lactone reductase activity exists in the extract of Mycobacterium bovis BCG cells [1]
Metabolism: It is oxidized by Mycobacterium tuberculosis L-gulonic acid-1,4-lactone dehydrogenase (Rv1771) to generate L-ascorbic acid (vitamin C) [1] |
| References | |
| Additional Infomation |
L-gulonic acid-1,4-lactone is the furanose form of gulonic acid lactone, with an L configuration. It is a metabolite in both humans and mice. It is functionally related to L-gulonic acid.
L-gulonic acid lactone has been reported to exist in Homo sapiens and Caenorhabditis elegans, and there is relevant data. L-gulonic acid-1,4-lactone is a metabolite found or produced in Saccharomyces cerevisiae. L-gulonic acid-1,4-lactone is a direct precursor of L-ascorbic acid (vitamin C) in animals, plants and some protists[1]. It is a highly specific substrate of Mycobacterium tuberculosis Rv1771 protein (L-gulonic acid-1,4-lactone dehydrogenase), which was the first reported bacterial enzyme specifically involved in vitamin C biosynthesis[1]. |
| Molecular Formula |
C6H10O6
|
|---|---|
| Molecular Weight |
178.1400
|
| Exact Mass |
178.047
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| CAS # |
1128-23-0
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| PubChem CID |
439373
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| Appearance |
White to off-white solid powder
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| Density |
1.8±0.1 g/cm3
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| Boiling Point |
467.9±18.0 °C at 760 mmHg
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| Melting Point |
187-190ºC(lit.)
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| Flash Point |
201.5±14.7 °C
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| Vapour Pressure |
0.0±2.6 mmHg at 25°C
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| Index of Refraction |
1.625
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| LogP |
-3.15
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
12
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| Complexity |
181
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| Defined Atom Stereocenter Count |
4
|
| SMILES |
C([C@@H]([C@@H]1[C@@H]([C@@H](C(=O)O1)O)O)O)O
|
| InChi Key |
SXZYCXMUPBBULW-SKNVOMKLSA-N
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| InChi Code |
InChI=1S/C6H10O6/c7-1-2(8)5-3(9)4(10)6(11)12-5/h2-5,7-10H,1H2/t2-,3+,4-,5+/m0/s1
|
| Chemical Name |
(3S,4R,5R)-5-[(1S)-1,2-dihydroxyethyl]-3,4-dihydroxyoxolan-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) |
H2O : ~50 mg/mL (~280.68 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.6136 mL | 28.0678 mL | 56.1356 mL | |
| 5 mM | 1.1227 mL | 5.6136 mL | 11.2271 mL | |
| 10 mM | 0.5614 mL | 2.8068 mL | 5.6136 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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