An alpha2,6-sialyltransferase cloned from Photobacterium leiognathi strain JT-SHIZ-119 shows both sialyltransferase and neuraminidase activity.
We cloned, expressed, and characterized a novel beta-galactoside alpha2,6-sialyltransferase from Photobacterium leiognathi strain JT-SHIZ-119. The protein showed 56-96% identity to the marine bacterial alpha2,6-sialyltransferases classified into glycosyltransferase family 80. The sialyltransferase activity of the N-terminal truncated form of the recombinant enzyme was 1477 U/L of Escherichia coli culture. The truncated recombinant enzyme was purified as a single band by sodium dodecyl sulfate polyacrylamide gel electrophoresis through 3 column chromatography steps. The enzyme had distinct activity compared with known marine bacterial alpha2,6-sialyltransferases. Although alpha2,6-sialyltransferases cloned from marine bacteria, such as Photobacterium damselae strain JT0160, P. leiognathi strain JT-SHIZ-145, and Photobacterium sp. strain JT-ISH-224, show only alpha2,6-sialyltransferase activity, the recombinant enzyme cloned from P. leiognathi strain JT-SHIZ-119 showed both alpha2,6-sialyltransferase and alpha2,6-linkage-specific neuraminidase activity. Our results provide important information toward a comprehensive understanding of the bacterial sialyltransferases belonging to the group 80 glycosyltransferase family in the CAZy database.
Mine
Toshiki
T
Glycotechnology Business Unit, Japan Tobacco Inc., Iwata, Shizuoka 438-0802, Japan.
Katayama
Sakurako
S
Kajiwara
Hitomi
H
Tsunashima
Masako
M
Tsukamoto
Hiroshi
H
Takakura
Yoshimitsu
Y
Yamamoto
Takeshi
T
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D010776:Photobacterium / Q000201:enzymology*; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012799:Sialyltransferases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D011994:Recombinant Proteins; D012799:Sialyltransferases; D009439:Neuraminidase; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase