Neisseria meningitidis is one type of bacteria responsible for causing bacterial meningitis. Based on the composition of sugar within its capsule, Neisseria meningitidis is classified into different serogroups. Six cause disease: serogroups A, B, C, X, Y and W. This study focuses on the N. meningitidis serogroup W capsule polymerase. This enzyme makes a capsular sugar containing galactose and sialic acid. Understanding the mechanism of this enzyme offers advantages in improving vaccine development against this serogroup. A simple and robust enzyme activity assay is essential to this knowledge. The overall goal of this work is to develop and optimize a fluorescence-based assay to monitor activity of this enzyme. Two different acceptors, DMB (1,2-diamino-4,5 methylenedioxybenzene) labeled sialic acid trimer and 4MUlac (4-methylumbelliferyl-lactoside) were used to test activity. These acceptors were first tested with a simple sialyltransferase from Camplyobacter jejuni, CSTII to assess activity. After these studies, the acceptors were tested with the serogroup W capsule polymerase. Results were visualized by TLC and HPLC with fluorescence detection. CST II enzyme showed activity with DMB trimer by both TLC and HPLC studies. The activity of the serogroup W enzyme with DMB trimer was observed by TLC but not by HPLC. In the case of 4 MUlac, no activity was observed with either enzyme. Studies are ongoing to test the activity of these enzymes with DMB labeled oligomers as well as 4MUlac again. Future studies will be focused on optimization once a suitable acceptor is determined.

Developing an HPLC based fluorescent assay for Neisseria meningitidis serogrouop W capsule polymerase

254th National Meeting (2017)

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