RESUMO
A system for production of human interferon-alpha2a (IFN-alpha2a) and IFN-alpha2b lacking N-terminal methionine has been developed. Plasmids containing genes of hybrid IFN-alpha2 under the control of different promoters were constructed; a sequence encoding the enteropeptidase hydrolysis site being introduced in proximal part of the genes. As the result, 4 strains of Escherichia coli producing hybrid IFN-alpha2 have been obtained. The methodology for IFN-alpha2 renaturation, hydrolysis of its N-terminal part, chromatographic purification of N-terminal methionine-free IFN-alpha2 has been developed.
Assuntos
Enteropeptidase/química , Microbiologia Industrial/métodos , Interferon-alfa/química , Interferon-alfa/isolamento & purificação , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Escherichia coli/enzimologia , Escherichia coli/genética , Vetores Genéticos/genética , Humanos , Interferon alfa-2 , Interferon-alfa/biossíntese , Metionina/química , Plasmídeos/genética , Renaturação Proteica , Proteínas Recombinantes de Fusão/biossíntese , Proteínas RecombinantesRESUMO
The effect of the composition of the nutrient medium on the biosynthesis of biotin by the culture Actinomyces species 313-152 was investigated. This microorganism can synthesize biotin on simple and complex media. The biotin synthesis is stimulated by oleic acid and Tween-80 utilized by the culture as a carbon source. The exogenous biotin added at concentrations of 1 x 10(-8) M to 1 x 10(-6) M to the medium containing fodder yeast hydrolyzate and Tween-80 did not inhibit the synthesis by the culture Actinomyces species 313-152.