114375-68-7Relevant articles and documents
Generation of a mutant mucor hiemalis endoglycosidase that acts on core-fucosylated N-Glycans
Katoh, Toshihiko,Katayama, Takane,Tomabechi, Yusuke,Nishikawa, Yoshihide,Kumada, Jyunichi,Matsuzaki, Yuji,Yamamoto, Kenji
, p. 23305 - 23317 (2016)
Endo-?-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-?1,6-fucosylated glycans. Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. Such mutant enzymes exhibited increased hydrolysis of a synthetic ?1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glycooxazoline) onto an ?1,6-fucosyl-N-acetylglucosaminyl biotin. Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics.
SUGAR COMPOUND, METHOD FOR PRODUCING SUGAR COMPOUND, ENGase ACTIVITY DETECTION COMPOSITION, NOVEL ENGase SCREENING METHOD, AND ENGase ACTIVITY INHIBITOR SCREENING METHOD
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Paragraph 0048, (2020/05/05)
PROBLEM TO BE SOLVED: To provide a compound for endo-β-N-acetylglucosaminidase activity detection. SOLUTION: The present invention provides a compound expressed by a formula I in which a combination of a fluorescent group and an extinction group causing fluorescence resonance energy transfer (FRET) is introduced into Z1 and Z2 [R, R' independently represent H or the like, R independently represent a single bond or the like]. SELECTED DRAWING: None COPYRIGHT: (C)2020,JPOandINPIT
