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Characterization of a chitinase gene (chiA) from Serratia marcescens BJL200 and one-step purification of the gene product
Authors:May B Brurberg  Vincent GH Eijsink  Ingolf F Nes
Institution:Norwegian Agricultural University, Laboratory of Microbial Gene Technology, P.O. Box 5051, 1432 Ås, Norway; The Norwegian State Agricultural Research Stations, 1432 Ås, Norway
Abstract:Abstract The nucleotide sequence of the chiA gene from Serratia marcescens strain BJL200 was determined. The gene was found to encode a protein of 563 amino acid residues, with a typical N-terminal signal peptide of 23 residues, that is cleaved off during export. The gene exhibited striking differences with two previously characterized chiA genes of S. marcescens in the region corresponding to amino acid residues 410–467 of the gene product. Periplasmic fractions of an Escherichia coli strain harbouring the cloned gene were used as starting material for the development of a fast, one-step purification protocol for the chitinase that is based on hydrophobic interaction chromatography.
Keywords:Serratia marcescens            Chitinase  Hydrophobic interaction chromatography
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