Purification and properties of glucosyltransferase responsible for water-insoluble glucan synthesis from Streptococcus mutans.

A glucosyltransferase responsible for water-insoluble glucan synthesis was purified from the culture fluids of Streptococcus mutans 6715-15 strain by column chromatography on Toyopearl HW-60 and subsequently on hydroxyapatite. The enzyme preparation gave a single band on analysis by polyacrylamide gel electrophoresis. The pH dependency of the activity showed two optimal peaks at 5.8 and 7.3 and the Km values for sucrose were 1.4 and 3.3 mM at the respective optimal pHs. The molecular weight determined by sodium dodecyl sulfate gel electrophoresis was 180,000. Although the enzyme scarcely synthesized water-insoluble and water-soluble glucans from sucrose, water-insoluble glucan formed from sucrose in the presence of dextran T10 consisted of over 93% alpha-1, 3-glucosidic linkage. Analysis of the structure of water-insoluble glucan indicated that the enzyme catalyzed the formation of branch points in alpha-1,6-glucan (dextran) and transferred the glucosyl moiety of sucrose to the C-3 position of the branching glucose residue of dextran. Since this enzyme has not yet been registered, we named it mutansynthetase (EC 2.4.1.?).