Chemo-enzymatic addition of a high-mannose type oligosaccharide to eel calcitonin (CT), a calcium-regulating hormone, was examined. The endo-β-N- acetylglucosaminidase from Arthrobacter protophormiae (Endo-A) transglycosylated the Man6-GlcNAc moiety from an ovalbumin-derived high- mannose type glycosyl asparagine, Asn(Man6-GlcNAc2)-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(Man6-GlcNAc2)3]-CT, was synthesized. The optimal reaction conditions for the synthesis of [Asn(Man6-GlcNAc2)3]-CT from Asn(Man6-GlcNAc2)-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(Man6-GlcNAc2)3]-CT was prepared.
All Science Journal Classification (ASJC) codes
- Applied Microbiology and Biotechnology