Vibrio vulnificus is the causative agent for the largest number of seafood borne fatalities in North America. Capsular polysaccharides (CPSs) that encapsulate the organism aids virulence. CPS biosynthesis has therefore become an attractive area of study due to the potential of targeting CPS production as an antimicrobial strategy. The biosynthesis of a CPS repeating unit from the clinical MO6-24 strain of Vibrio vulnificus requires three general enzyme types: sugar-modifying enzymes, glycolsyltransferases, and a hexose-1-phosphate transferase. The hexose-1-phosphate transferase is responsible for the first sugar transfer onto an isoprenoid membrane anchor. We have identified this hexose-1-phosphate transferase to be the protein encoded by the CPS biosynthesis gene wbfU. WbfU was expressed and purified, then using HPLC methods the activity of the protein was assayed with a fluorescently labeled isoprenoid. Surprisingly, we found that this protein is capable of transferring several types of sugars and provides an excellent enzymatic tool for the construction of a variety of isoprenoid-linked monosaccharides. By identifying the first step in capsule biosynthesis we expect to be able to characterize the remaining steps in V. vulnificus capsule production in the first in vitro reconstitution of the biosynthesis of this complex polymer.

Characterization of the hexose-1-phosphate transferase from the Vibrio vulnificus capsular polysaccharide biosynthesis pathway

249th National Meeting (2015)

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