Purification and characterization of a marine bacterial beta-galactoside alpha 2,6-sialyltransferase from Photobacterium damsela JT0160.
A bacterial sialyltransferase, named sialyltransferase 0160, was purified from a marine bacterium that had been isolated from seawater from Sagami Bay, Kanagawa. This strain has been identified as Photobacterium damsela, and named P. damsela JT0160. Sialyltransferase 0160 was purified 688-fold to homogeneity from the crude extract of the cells with a yield of 19% using a combination of anion exchange chromatography, hydroxyapatite chromatography, gel filtration chromatography, and affinity chromatography. The purified enzyme migrated as a single band (61 kDa) on sodium dodecyl sulfate-polyacrylamide gel. This sialyltransferase was found to be a beta-galactoside alpha 2,6-sialyltransferase [EC 2.4.99.1] which catalyzes the incorporation of NeuAc from CMP-NeuAc into the galactose residue of the carbohydrate chain at position 6 on the basis of an analysis of the enzymatic reaction products with HPLC, 1H-, 13C-NMR spectroscopy, and fast atom bombardment mass spectroscopy.
Yamamoto
T
T
Sea Water Science Research Laboratory, Japan Tobacco Inc., Kanagawa.
Nakashizuka
M
M
Kodama
H
H
Kajihara
Y
Y
Terada
I
I
D002240:Carbohydrate Sequence; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007785:Lactose / Q000378:metabolism; D008756:Methylgalactosides / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010776:Photobacterium / Q000201:enzymology*; D012799:Sialyltransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D008756:Methylgalactosides; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid; D012799:Sialyltransferases; D007785:Lactose; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase