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A novel amino acid supplementation strategy based on a stoichiometric model to enhance human IL-2 (interleukin-2) expression in high-cell-density Escherichia coli cultures.
- Source :
-
Biotechnology and applied biochemistry [Biotechnol Appl Biochem] 2010 Dec; Vol. 57 (4), pp. 151-6. - Publication Year :
- 2010
-
Abstract
- A novel amino acid supplementation strategy was developed for enhancing the production of IL-2 (interleukin-2; as a model protein) by recombinant Escherichia coli BL21 (pET21a-hil2) in fed-batch high-cell-density cultures. The amino acids most needed and their amounts were determined using a stoichiometric model, and full factorial design experiments were conducted to determine the effects of single amino acids and amino acid mixtures on production. One of the most effective amino acid mixtures was found to be leucine, aspartic acid and glycine. This amino acid mixture was utilized for the production of IL-2 in batch and fed-batch fermentations. The amount of IL-2 produced increased from 403 to 722 mg/l and from 5.15 × 10³ to 8.08 × 10³ mg/l in batch and fed-batch cultures respectively. The results also revealed that the above amino acid mixture specifically increases IL-2 concentration in the cells.
- Subjects :
- Acetates metabolism
Culture Media
Escherichia coli genetics
Escherichia coli growth & development
Fermentation
Glucose metabolism
Humans
Interleukin-2 chemistry
Interleukin-2 genetics
Kinetics
Models, Molecular
Recombinant Proteins chemistry
Recombinant Proteins genetics
Amino Acids metabolism
Escherichia coli metabolism
Interleukin-2 biosynthesis
Recombinant Proteins biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1470-8744
- Volume :
- 57
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Biotechnology and applied biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 21062264
- Full Text :
- https://doi.org/10.1042/BA20100320