Cloning, functional expression and partial characterization of the glucosekinase from Renibacterium salmoninarum

The complete glcK gene from the fish pathogen Renibacterium salmoninarum, e ncoding a glucose kinase, was analyzed and expressed. The partial character ization of the recombinant enzyme confirmed that it belongs to a group of g lucose kinases involved in carbon catabolite repression. Multiple sequence alignments were used to deduce a new consensus sequence for this family of bacterial proteins, characterized by several conserved Cys residues. This s equence was more specific and allowed the detection of the first eukaryotic protein of this family. The recombinant enzyme was inhibited by N-ethylmal eimide and the substrates protected the enzyme from this inhibition, sugges ting the presence of Cys residues in or close to the active site. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Sci ence B.V. All rights reserved.