Cloning and characterization of the gene encoding 3-deoxy-D-manno-octulosonate 8-phosphate synthetase from Escherichia coli

Abstract

The cloning of the gene for Escherichia coli PL-2 2-keto-3-deoxy-D-manno-octonate 8-phosphate synthetase is reported. Positive transformants showed an increase of approximately three-fold in specific activity of the enzyme both in E. coli and in Salmonella typhimurium as host cells. A subclone containing a 1.5-kilobase PvuII fragment overproduced active enzyme. Minicell experiments that allow the detection of plasmid encoded proteins revealed an insert-coded single protein band of 34 kilodaltons.