Physicochemical Properties
| CAS # | 9028-25-5 |
| Appearance | Typically exists as solid at room temperature |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets | The target of myo-Inositol dehydrogenase is myo-inositol (a cyclic polyol), which catalyzes the NAD⁺-dependent dehydrogenation of myo-inositol. |
| ln Vitro |
1. Heterologous expression and purification of myo-Inositol dehydrogenase from Lactobacillus casei BL23 were performed in Escherichia coli. The purified enzyme showed catalytic activity towards myo-inositol, with NAD⁺ serving as the cofactor. [1] 2. X-ray crystallography analysis revealed that myo-Inositol dehydrogenase binds its substrate myo-inositol in a specific orientation distinct from that of scyllo-inositol dehydrogenase (another inositol dehydrogenase from the same strain). The substrate binding pocket of myo-Inositol dehydrogenase has unique amino acid residues (e.g., Tyr155, Asp220) that mediate interactions with myo-inositol, contributing to its substrate specificity. [1] 3. Enzymatic activity assays confirmed that myo-Inositol dehydrogenase exhibits higher specificity for myo-inositol compared to other inositol isomers (e.g., scyllo-inositol, D-chiro-inositol), as reflected by differences in catalytic efficiency (kcat/Km). [1] |
| Enzyme Assay |
1. myo-Inositol dehydrogenase activity was measured by monitoring the reduction of NAD⁺ to NADH (detected at 340 nm, an absorbance peak specific to NADH) in a reaction system. The standard reaction mixture contained a buffer (e.g., Tris-HCl buffer, pH 8.0), NAD⁺ (at a final concentration of 1 mM), myo-inositol (at varying concentrations to determine kinetic parameters), and purified myo-Inositol dehydrogenase. The reaction was initiated by adding the enzyme, and the change in absorbance at 340 nm was recorded continuously for a certain period (e.g., 5-10 minutes) at 37°C. Kinetic parameters (Km for myo-inositol and NAD⁺, kcat) were calculated by fitting the initial velocity data to the Michaelis-Menten equation. [1] 2. For analyzing substrate specificity, the above assay was repeated by replacing myo-inositol with other inositol isomers (e.g., scyllo-inositol, D-chiro-inositol) at the same concentration, and the relative enzymatic activity was compared to that with myo-inositol. [1] 3. To study the effect of pH on enzyme activity, the reaction buffer was adjusted to different pH values (e.g., pH 6.0-9.0 using citrate-phosphate buffer, Tris-HCl buffer, or glycine-NaOH buffer), and the enzymatic activity was measured as described above to determine the optimal pH for myo-Inositol dehydrogenase. [1] |
| References |
[1]. Aamudalapalli HB, et.al. myo-Inositol dehydrogenase and scyllo-inositol dehydrogenase from Lactobacillus casei BL23 bind their substrates in very different orientations. Biochim Biophys Acta Proteins Proteom. 2018 Nov;1866(11):1115-1124. |
| Additional Infomation |
1. myo-Inositol dehydrogenase is a member of the short-chain dehydrogenase/reductase (SDR) superfamily, characterized by a conserved Rossmann fold (a structural motif involved in cofactor binding). [1] 2. The distinct substrate binding orientation of myo-Inositol dehydrogenase (compared to scyllo-inositol dehydrogenase) is crucial for its role in the inositol metabolic pathway of Lactobacillus casei BL23, which allows the strain to utilize myo-inositol as a carbon source. [1] 3. Site-directed mutagenesis experiments on key amino acid residues in the substrate binding pocket (e.g., Tyr155Ala, Asp220Ala) resulted in a significant decrease in myo-Inositol dehydrogenase activity, confirming the importance of these residues in substrate binding and catalysis. [1] |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |