Purification and characterization of glucosyltransferases from Streptococcus mutans 6715.
A water-soluble glucan-synthesizing glucosyltransferase (GTase-S) and a water-insoluble glucan-synthesizing glucosyltransferase (GTase-I) were purified from culture supernatant of Streptococcus mutans 6715 (serotype g) by ammonium sulphate precipitation, chromatofocusing on a Polybuffer exchanger PBE 94 column, and subsequent phenyl-Sepharose CL-4B or hydroxyapatite column chromatography. The GTase-S and GTase-I activities were purified 4019- and 4714-fold, respectively, and the molecular weights were calculated to be 160000 and 165000, respectively. GTase-S had a pH optimum of 5.0, a Km of 8.8 mM for sucrose in the presence of 20 microM-dextran T10, and an isoelectric point of pH 4.3. GTase-I had two pH optima of 5.0 and 7.0, Km values of 4.9 mM (at pH 5.0) and 7.0 mM (at pH 7.0), mM (at pH 7.0), and an isoelectric point of pH 4.9. Methylation analysis indicated that the water-soluble glucan produced by GTase-S was a highly branched 1,6-alpha-linked D-glucan with 1,3-linked glucose residues, and that the water-insoluble glucan synthesized by GTase-I was composed of 1,3-alpha-linked glucose units.
Furuta
T
T
Koga
T
T
Nisizawa
T
T
Okahashi
N
N
Hamada
S
S
D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D002845:Chromatography; D003911:Dextrans / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005936:Glucans / Q000096:biosynthesis; D005964:Glucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D007525:Isoelectric Focusing; D013295:Streptococcus mutans / Q000201:enzymology*
D003911:Dextrans; D005936:Glucans; D005964:Glucosyltransferases; D000645:Ammonium Sulfate