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

Автор: 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, 1540 просмотров.