Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli
| Autor: | N.A. Terenteva, L.A. Balabanova, V.A. Golotin, N.S. Vorobieva, L.V. Slepchenko, Yu. A. Noskova, I. Yu Bakunina, Valery A. Rasskazov |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Erythrocytes CAZy Acclimatization Cell Culture Techniques lac operon medicine.disease_cause 01 natural sciences law.invention Microbiology 03 medical and health sciences Pseudoalteromonas law 010608 biotechnology Escherichia coli medicine Humans Cloning Molecular Alpha-galactosidase biology Strain (chemistry) biology.organism_classification Recombinant Proteins Cold Temperature 030104 developmental biology Biochemistry alpha-Galactosidase Recombinant DNA biology.protein Heat-Shock Response Bacteria Biotechnology |
| Zdroj: | Europe PubMed Central |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2016.03.006 |
| Popis: | α-Galactosidase (α-PsGal) of the cold-adapted marine bacterium Pseudoalteromonas sp. KMM 701 was cloned into the pET-40b(+) vector to study its properties and to develop an effective method for modifying human B-erythrocytes into O-blood group. The use of heat-shock as a pre-induction treatment, IPTG concentration of 0.2 mM and post-induction cultivation at 18 °C for 20 h in the developed MX-medium allowed increasing the recombinant Escherichia coli Rosetta (DE3)/40Gal strain productivity up to 30 times and the total soluble α-PsGal yield up to 40 times. |
| Databáze: | OpenAIRE |
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