Partial purification and properties of UDP-N-acetylmannosamine:N-acetylglucosaminyl pyrophosphorylundecaprenol N-acetylmannosaminyltransferase from Bacillus subtilis.
An enzyme which catalyzes the conversion of GlcNAc-PP-undecaprenol into ManNAc(beta 1----4)GlcNAc-PP-undecaprenol, a key lipid intermediate in the de novo synthesis of various teichoic acids, was partially purified from the 20,000 x g supernatant fraction of Bacillus subtilis AHU 1035 cell homogenate. By means of ammonium sulfate precipitation, gel chromatography, and ion-exchange chromatography, the enzyme was purified about 70-fold, giving a preparation virtually free from substances obstructive to measurement of the N-acetylmannosaminyltransferase reaction. The enzyme was shown to be specific to UDP-ManNAc. The Km value for UDP-ManNAc was 4.4 microM, and the optimum pH was 7.3. The enzyme required 10 mM MgCl2, 0.3 M KCl, 25% glycerol, and 0.1% Nonidet P-40 to function at full activity.
Murazumi
N
N
Department of Chemistry, Faculty of Science, Hokkaido University.
Kumita
K
K
Araki
Y
Y
Ito
E
E
D001412:Bacillus subtilis / Q000201:enzymology*; D002236:Carbohydrate Conformation; D002845:Chromatography; D005964:Glucosyltransferases; D006017:Glycolipids; D006602:Hexosyltransferases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008364:Mannosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases; D017349:N-Acetylhexosaminyltransferases; D013379:Substrate Specificity; D013682:Teichoic Acids / Q000096:biosynthesis; D000637:Transaminases / Q000302:isolation & purification*; D014539:Uridine Diphosphate Sugars
D006017:Glycolipids; D013682:Teichoic Acids; D014539:Uridine Diphosphate Sugars; C052604:N-acetylglucosamine-pyrophosphorylundecaprenol; C059191:uridine diphosphate N-acetylmannosamine; D005964:Glucosyltransferases; D006602:Hexosyltransferases; D008364:Mannosyltransferases; D017351:N-Acetylglucosaminyltransferases; D017349:N-Acetylhexosaminyltransferases; C059053:N-acetylmannosaminyltransferase; C059192:UDP-N-acetylmannosamine-N-acetylglucosaminyl pyrophosphorylundecaprenol N-acetylmannosaminyltransferase; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I; D000637:Transaminases