L-Alanyl-L-GLutamine ReadyMade™ Solution (syn: Ala-Gln) is a sterile-filtered solution of L-Alanyl-L-Glutamine dipeptide in 0.85% sodium chloride (NaCl) at a concentration of 200 mM.
This product is an alternative to L-Glutamine, an amino acid needed by mammalian cells. L-Glutamine is more unstable in cell culture solutions compared to other amino acids, and the rate of degradation of L-Glutamine depends on storage temperature, product age, and pH. Ala-Gln has better stability compared to L-Glutamine alone while in solution and is also referred to as 'stable glutamine. It is suitable for mammalian cell culture and biopharmaceutical applications.
Note: The formulation contains 43.44 mg/ml of L-alanyl-L-glutamine in 0.85% NaCl.
The working concentration for Ala-Gln is typically 1 to 4 mM. Thus 10 ml is sufficient for up to one liter of cell culture medium. Each user should determine the optimal working concentration for their application.
| Mechanism of Action |
Ala-Gln is hydrolyzed to release glutamine and alanine in vivo. While it acts as a source of glutamine, Ala-Gln is used as a substitute for L-glutamine both in vitro and in vivo. While L-glutamine can generate ammonia when added to cell cultures grown at 37°C which can cell growth, Ala-Gln causes a reduction in ammonia production, and thereby can reduce the apoptotic ratio of cells grown in culture, leading to beneficial effects. |
| Eukaryotic Cell Culture Applications |
Ala-Gln does not spontaneously break down to form ammonia. Instead, cells cleave the dipeptide bond in the molecule to release L-Glutamine, an essential amino acid and required additive for most cell culture media. This means that the cell cultures have a lower level of ammonia which improves their viability. In addition, the product extends the shelf life at 4-8°C and is stable across a wider temperature range compared to L-Glutamine alone. Ala-Gln is used a supplement in Dulbecco′s Modified Eagle Medium (DMEM) media, a popular medium to culture human cell lines. In fact, Ala-Gln is the preferred L-glutamine source for primary cell cultures, stem cells, and high metabolic cells. BJECs were exposed (or not) to 1 μg/mL LPS for 24 h to generate a pro-inflammatory model. The cells were then treated with different concentrations of Ala-Gln (0.25, 0.5, 1.0, 2.0, or 4.0 mmol/L) to detect any regulatory effects on the inflammation and barrier function of BJECs. L-GLutamine is involved in the formation of purine and pyrimidine nucleotides, amino sugars, glutathione, L-glutamate, and other amino acids, and is also involved in protein synthesis and glucose production. Intestinal porcine epithelial cells (IPEC-J2) were cultured for 2 days in Dulbecco’s modified Eagle’s-F12 Ham medium (DMEM-F12) with 2.5 mM (L-glutamine), Ala-Gln or glycyl-glutamine (Gly-Gln). There were no differences in proliferation between free L-Glutamine and Ala-Gln-treated cells. Ala-Gln treatment increased the basal respiration and ATP production, compared with free L-Glutamine. Authors concluded that that Ala-Gln has a very similar functional profile to free L-Glutamine in porcine enterocytes in vitro (Tan et al, 2017). Researchers investigated the effects Ala-Gln on the regulation of lipopolysaccharide (LPS)-induced inflammation and barrier function in bovine jejunum epithelial cells (BJECs). It rescued the decrease in cell viability and prevented the accumulation of ILs after LPS exposure by reducing the mRNA and protein expression levels of inflammatory factors. In conclusion, Ala-Gln attenuated injury from inflammation and repaired damaged intestinal barrier induced with LPS (Zhang et al, 2019). |
| Molecular Formula | C8H15N3O4 |
| References |
Hassell T, Gleave S, Butler M (1991) Growth inhibition in animal cell culture. The effect of lactate and ammonia. Appl Biochem. Biotechnol. 30(1):29-41 PMID 1952924 Tan B, Liu H, He G et al (2017) Alanyl-glutamine but not glycyl-glutamine improved the proliferation of enterocytes as glutamine substitution in vitro. Amino Acids 49, 2023–2031 Zhang X et al (2019) Alanyl-glutamine ameliorates lipopolysaccharide-induced inflammation and barrier function injury in bovine jejunum epithelial cells. Biochem. and Cell Biol 97(6): 670-680 Zhu J, Yang W, Wang B et al (2020) Metabolic engineering of Escherichia coli for efficient production of l-alanyl-l-glutamine. Microb. Cell Fact .19:129 |
