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Development of Industrial Technology for Cultivation of E. coli Strain Producing an Enzyme of ULP-Proteinase that Accomplishes Proteolytic Processing of SUMO-IFN, a Hybrid Protein Precursor of a-Interferon
Development of Industrial Technology for Cultivation of E. coli Strain Producing an Enzyme of ULP-Proteinase that Accomplishes Proteolytic Processing of SUMO-IFN, a Hybrid Protein Precursor of a-InterferonАвтор: N.A. Litvinova, N.V. Stratonova, V.S. Leonov, and R.A. Khamitov Страница: 60-70
Development of Industrial Technology for Cultivation of E. coli Strain Producing an Enzyme of ULP-Proteinase that Accomplishes Proteolytic Processing of SUMO-IFN, a Hybrid Protein Precursor of a-Interferon Russian Journal of Biotechnology, 2013, N 5, P. 60-70 UDC 604.4:577.15 Section: “Biologicals Technology”
N.A. Litvinova *, N.V. Stratonova, V.S. Leonov, and R.A. Khamitov The State Research Institute for Genetics and Selection of Industrial Microorganisms, 117545, Moscow Russia e-mail: natalya.litvinova@bioprocess.ru
A method for the industrial cultivation of an E. coli strain producing ULP-proteinase has been developed. The composition of nutrient medium, process of launching of the induction of the target protein expression and mass-exchange characteristic of the cultivation were optimized. The growing should be performed at 20°C without medium dilution; lactose (0.5%) should be used as an inducer; pH should be 6.9—7.1; and aeration should be equal to 10 l/min at mixing of 300 rpm. The productivity by the target protein achieved under the above conditions was (32±0.1) mg/l of CL, whereas the chromatography-purified preparation of ULP-proteinase had more than 95% purity.
Key word: hybrid precursor protein, interferon alpha-2b, optimization, SUMO, ULP-proteinase.
01.04.2015, 1610 просмотров. |
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