| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Butyryl Coenzyme A serves as a substrate for Butyryl-CoA:acetate-CoA transferase (But, EC 2.8.3.8). This enzyme catalyzes the transfer of the CoA group from butyryl-CoA to acetate. [1]
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| ln Vitro |
- The activity of butyryl-CoA:acetate-CoA transferase (But) was measured using a citrate synthase-coupled assay. The enzyme uses butyryl-CoA and acetate as substrates. The generated acetyl-CoA is condensed with oxaloacetate by citrate synthase, liberating free CoA, which reacts with 5,5'-dithio-bis-(2-nitrobenzoate) to form a yellow thiophenolate anion. Reaction rates were monitored by measuring absorbance at 412 nm at 39°C. [1]
- Among 12 putative but genes identified from swine intestinal butyrate-producing bacterial strains, eight genes encoded proteins with strong But enzyme activity, with specific activities ranging from 7,004 μM·min⁻¹·mg⁻¹ (strain 27-5-10) to 27,819 μM·min⁻¹·mg⁻¹ (strain 831b). When propionyl-CoA was used as a substrate, these highly active genes exhibited similar activity. [1] - Highly active But proteins contain a conserved amino acid motif LQLGIGG. The glycine residue in this motif was conserved in all highly active sequences, while proteins with low But activity contained at least one substitution in this motif. [1] |
| ln Vivo |
- In swine proximal colon contents, RNA-based but gene libraries revealed greater OTU diversity than DNA-based libraries (Shannon diversity index with Wilcoxon paired test, P < 0.03; Shannon evenness with Wilcoxon paired test, P < 0.03), indicating that the actively transcribing butyrate-producing community differs from the most abundant members. [1]
- A total of 92 unique but gene OTUs (97% similarity) were detected from DNA and RNA extracted from the colonic contents of six pigs. Fourteen OTUs were detected in RNA libraries from every pig, four OTUs were detected in all DNA libraries, and three OTUs (OTU4, OTU14, OTU23) were detected in every library regardless of nucleic acid type. [1] |
| Enzyme Assay |
- Citrate synthase-coupled assay for butyryl-CoA transferase activity: The reaction system contains the substrates butyryl-CoA and acetate. The butyryl-CoA:acetate-CoA transferase catalyzes the transfer of CoA, producing butyrate and acetyl-CoA. The generated acetyl-CoA is condensed with oxaloacetate by citrate synthase, releasing free CoA. The released CoA reacts with 5,5'-dithio-bis-(2-nitrobenzoate) to form a yellow thiophenolate anion. Reaction rates were measured by monitoring absorbance at 412 nm at 39°C. Crude cell lysates were used for the assay and diluted with sterile water as necessary to achieve the linear range for the reaction rate. The reaction was repeated in the absence of acetate to confirm that the measured rate was not due to CoA-hydrolase activity. [1]
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| Cell Assay |
- Gene cloning and protein expression: Candidate but genes were cloned into the pET-TOPO-101 vector and transformed into TOP10 E. coli chemically competent cells. Positive clones with full-length gene inserts were confirmed by sequencing. Cloned DNAs were transformed into E. coli BL21 Star competent cells for protein expression. Cultures (100 ml) were grown for 12 h in LB containing 50 μg/ml carbenicillin. Expression was induced by adding isopropyl-β-D-thiogalactopyranoside (IPTG) to a final concentration of 1 mM. After an additional 6 h of growth, cultures were harvested by centrifugation, washed, and resuspended in sterile phosphate-buffered saline (PBS). Cells were lysed by two passages through a French press. Lysates were centrifuged to remove remaining unlysed cells, and the supernatant was used for activity assays. [1]
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| Animal Protocol |
- Sample collection and processing: Proximal colon contents (10 cm distal from the cecum) from pigs were immediately placed in RNAlater and quickly homogenized to preserve nucleic acid integrity. Samples were subsequently frozen at -80°C until extraction (within 1 month). DNAs and RNAs were extracted from proximal colon contents of six pigs fed a standard diet using DNA and RNA extraction kits. The iScript Select kit was used to generate cDNA from the RNA using random hexamer primers. [1]
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| References | |
| Additional Infomation |
- Background: Butyrate is essential for colonic homeostasis and is the preferred energy source for colonocytes. Butyrate reduces local oxygen concentrations, causing epithelial hypoxia and limiting the growth of facultative aerobic pathogens like Salmonella. Additionally, butyrate alters host gene expression to promote immune tolerance to the colonic microbiota and improve colonic epithelial barrier function. [1]
- Butyrate synthesis pathway: The most common pathway for butyrate production in colonic environments involves the condensation of two molecules of acetyl-CoA, followed by reduction to butyryl-CoA. After butyryl-CoA is generated, two different enzymes are responsible for the final conversion to butyrate: butyrate kinase (Buk) and butyryl-CoA:acetate-CoA transferase (But), with the But protein being the most common in the colonic environment. [1] - Phylogenetic analysis: The genes encoding highly functional But proteins are phylogenetically separated from potential paralogues, but this separation is not absolute. The vast majority of OTUs detected by the funbut primers (99.4%) clustered with but sequences encoding highly active enzymes, while only 0.6% of sequences fell outside this main functional clade. [1] |
| Molecular Formula |
C25H39LI3N7O17P3S
|
|---|---|
| Molecular Weight |
855.423147439957
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| Exact Mass |
855.181610
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| CAS # |
102282-28-0
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| Related CAS # |
Butyryl-Coenzyme A lithium hydrate; Butyryl-Coenzyme A trisodium; Butyryl-Coenzyme A triammonium
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| Appearance |
Typically exists as solids at room temperature
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| SMILES |
S(C(CCC)=O)CCNC(CCNC([C@@H](C(C)(C)COP(=O)([O-])OP(=O)(O)OC[C@@H]1[C@H]([C@H]([C@H](N2C=NC3C(N)=NC=NC2=3)O1)O)OP(=O)([O-])[O-])O)=O)=O.[Li+].[Li+].[Li+]
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| Synonyms |
RefChem:571907; Butyryl coenzyme A lithium salt hydrate; 634-407-5; Butyryl Coenzyme A lithium salt; Butyryl CoA trilithium
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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) |
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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1690 mL | 5.8451 mL | 11.6902 mL | |
| 5 mM | 0.2338 mL | 1.1690 mL | 2.3380 mL | |
| 10 mM | 0.1169 mL | 0.5845 mL | 1.1690 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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