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1,2‐α‐ l ‐Fucosynthase: A glycosynthase derived from an inverting α‐glycosidase with an unusual reaction mechanism
Author(s) -
Wada Jun,
Honda Yuji,
Nagae Masamichi,
Kato Ryuichi,
Wakatsuki Soichi,
Katayama Takane,
Taniguchi Hajime,
Kumagai Hidehiko,
Kitaoka Motomitsu,
Yamamoto Kenji
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.09.054
Subject(s) - glycoside hydrolase , chemistry , asparagine , stereochemistry , beta galactosidase , lactose , mutant , enzyme , biochemistry , gene expression , gene
Fucosyloligosaccharides have great therapeutic potential. Here we present a new route for synthesizing a Fucα1,2Gal linkage by introducing glycosynthase technology into 1,2‐α‐ l ‐fucosidase. The enzyme adopts a unique reaction mechanism, in which asparagine‐423 activated by aspartic acid‐766 acts as a base while asparagine‐421 fixes both a catalytic water and glutamic acid‐566 (an acid) in the proper orientations. Glycosynthase activity of N421G, N423G, and D766G mutants was examined using β‐fucosyl fluoride and lactose, and among them, the D766G mutant most effectively synthesized 2′‐fucosyllactose. 1,2‐α‐ l ‐Fucosynthase is the first glycosynthase derived from an inverting α‐glycosidase and from a glycosidase with an unusual reaction mechanism.