title: Purification and characterization of cell-associated glucosyltransferase synthesizing insoluble glucan from Streptococcus mutans serotype c.
abstract: Streptococcus mutans Ingbritt (serotype c) was shown to have a significant amount of cell-associated glucosyltransferase activity which synthesizes water-insoluble glucan from sucrose. The enzyme was extracted from the washed cells with SDS, renatured with Triton X-100, adsorbed to 1,3-alpha-D-glucan gel, and then eluted with SDS. The enzyme preparation was electrophoretically homogeneous, and the specific activity was 7.3 i.u. (mg protein)-1. The enzyme had an Mr of 158,000 as determined by SDS-PAGE, and was a strongly hydrophilic protein, as judged by its amino acid composition. The enzyme gradually aggregated in the absence of SDS. The enzyme had an optimum pH of 6.5 and a Km value of 16.3 mm for sucrose. Activity was stimulated 1.7-fold by dextran T10, but was not stimulated by high concentrations of ammonium sulphate. Below a sodium phosphate buffer concentration of 50 mm, activity was reduced by 75%. This enzyme synthesized an insoluble D-glucan consisting of 76 mol% 1,3-alpha-linked glucose and 24 mol% 1,6-alpha-linked glucose.
authors.affiliation: Department of Chemistry, National Defense Medical College, Saitama, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Mukasa
authors.forename: A
authors.initials: A
authors.lastname: Shimamura
authors.forename: H
authors.initials: H
authors.lastname: Tsumori
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism; D005936:Glucans / Q000096:biosynthesis*; D005964:Glucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D012995:Solubility; D013295:Streptococcus mutans / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D005936:Glucans; C036655:1,3-alpha-D-glucan synthase; D005964:Glucosyltransferases