MOLECULAR-CLONING AND SEQUENCE-ANALYSIS OF FLAVASTACIN - AN O-GLYCOSYLATED PROKARYOTIC ZINC METALLOENDOPEPTIDASE

A new zinc metalloendopeptidase that cleaves peptides on the amino-ter minal side of aspartic acid was isolated from the cultural filtrate of Flavobacterium meningosepticum. The gene for this new enzyme was clon ed into pBluescript, and the complete nucleotide sequence was determin ed. Over 40% of the deduced amino acid sequence was verified independe ntly by direct protein microsequencing. The most important structural features of this new enzyme include (i) the presence of an unusual O-l inked oligosaccharide of unknown function located at a unique consensu s site near the C-terminus and (ii) a characteristic extended zinc-bin ding site and corresponding Met-turn that places this metalloendopepti dase in the astacin family. This is the first example of a prokaryotic enzyme related to the eukaryotic astacin group; it is being designate d hereafter as flavastacin. (C) 1995 Academic Press, Inc.