Purification, resolution, and interaction of the glucosyltransferases of Streptococcus mutans 6715.
The extracellular glucosyltransferase produced by Streptococcus mutans 6715 was purified from culture supernatant fluids to a specific activity of 9.6 IU/mg of protein, with an overall recovery of 87%. The purified enzyme preparation, designated unfractionated enzyme (UFE), synthesized only water-insoluble glucans from sucrose during the initial stages of the reaction, although some water-soluble polymers accumulated after extended periods of incubation. It was free from measurable fructosyltransferase activity. The UFE preparation was resolved into two different catalytically active components by ethanol fractionation. One fraction (designated insoluble product enzyme [IPE]) synthesized water-insoluble glucans, whereas the other (designated soluble product enzyme [SPE]) produced primarily water-soluble glucans. The difference between the insoluble glucans made by the UFE preparation and those made by the IPE fraction appeared to be due to interaction of the SPE and IPE components in the UFE preparation. Addition of commercial dextrans or enzymatically prepared glucans to the glucosyltransferase assay altered the amounts of soluble and insoluble glucans synthesized by the UFE preparation. The molecular weights of the major enzymatically active proteins producing insoluble and soluble glucans were estimated by gel filtration chromatography to be 150,000 and 175,000, respectively.
D003910:Dextranase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005964:Glucosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000378:metabolism; D011134:Polysaccharides / Q000096:biosynthesis; D012995:Solubility; D013295:Streptococcus mutans / Q000201:enzymology* / Q000378:metabolism; D013395:Sucrose / Q000378:metabolism
D006023:Glycoproteins; D009097:Multienzyme Complexes; D011134:Polysaccharides; D013395:Sucrose; D005964:Glucosyltransferases; D003910:Dextranase