Expression and rapid one-step purification of biologically active His-tagged factor C by Ni(2+) affinity column chromatography

• Published in: FEMS microbiology letters Vol. 196; no. 2; pp. 223 - 227
• Main Authors: Birkó, Z, Schauwecker, F, Pfennig, F, Szeszák, F, Vitális, S, Keller, U, Biró, S
• Format: Journal Article
• Language: English
• Published: England 15.03.2001
• Subjects: Amino Acid Sequence; Amino Acids - analysis; Bacterial Proteins - biosynthesis; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Base Sequence; Chromatography, Affinity; Cloning, Molecular; DNA, Bacterial; Escherichia coli; Genes, Bacterial; Histidine - metabolism; Molecular Sequence Data; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Streptomyces griseus - genetics; Streptomyces griseus - metabolism; Streptomyces griseus - ultrastructure; Transformation, Bacterial
• ISSN: 0378-1097
• DOI: 10.1016/S0378-1097(01)00072-6

Factor C is an unusual extracellular protein capable of inducing cytodifferentiation in certain Streptomyces strains. The protein is produced by Streptomyces griseus 45H at such a low amount that the study of its mode of action was hindered by the shortage of purified protein. We report here the expression of C-terminally hexa-His-tagged factor C in Streptomyces lividans and Escherichia coli. Expression in S. lividans is low while in E. coli it is relatively high, yielding about 5--10 mg of biologically fully active protein per liter culture.