title: Chemo-enzymatic synthesis of a calcitonin derivative containing a high-mannose type oligosaccharide by endo-beta-N-acetylglucosaminidase from Arthrobacter protophormiae.
abstract: Chemo-enzymatic addition of a high-mannose type oligosaccharide to eel calcitonin (CT), a calcium-regulating hormone, was examined. The endo-beta-N-acetylglucosaminidase from Arthrobacter protophormiae (Endo-A) transglycosylated the Man(6)-GlcNAc moiety from an ovalbumin-derived high-mannose type glycosyl asparagine, Asn(Man(6)-GlcNAc(2))-OH, to a calcitonin derivative, [Asn(GlcNAc)(3)]-CT, in which the N-acetyl-d-glucosamine (GlcNAc) is attached to the third l-asparagine (Asn) residue of the peptide, and a calcitonin derivative containing a high-mannose type oligosaccharide, [Asn(Man(6)-GlcNAc(2))(3)]-CT, was synthesized. The optimal reaction conditions for the synthesis of [Asn(Man(6)-GlcNAc(2))(3)]-CT from Asn(Man(6)-GlcNAc(2))-OH and [Asn(GlcNAc)(3)]-CT catalyzed by Endo-A were examined. The transglycosylation yield relative to the concentration of the [Asn(GlcNAc)(3)]-CT added was 32.7%, and 4.42 mg of [Asn(Man(6)-GlcNAc(2))(3)]-CT was prepared.
authors.affiliation: The Noguchi Institute, 1-8-1 Kaga, Itabashi, Tokyo 173-0003 Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: K
authors.initials: K
authors.lastname: Haneda
authors.forename: R
authors.initials: R
authors.lastname: Iguchi
authors.forename: T
authors.initials: T
authors.lastname: Inazu
authors.forename: M
authors.initials: M
authors.lastname: Mizuno
authors.forename: K
authors.initials: K
authors.lastname: Takegawa
authors.forename: A
authors.initials: A
authors.lastname: Kondo
authors.forename: I
authors.initials: I
authors.lastname: Kato