Chemo-enzymatic synthesis of glycosylated insulin using a GlcNAc tag.
Artificial insulin with an N-linked oligosaccharide was synthesized by a chemo-enzymatic method using endo-beta-N-acetylglucosaminidase from Mucor hiemalis (Endo-M). GlcNAc-modified insulin was prepared by the reaction of the carboxymethyl glycoside of GlcNAc and 3 amino groups of bovine insulin using a dimethylphosphinothioic mixed anhydride (Mpt-MA) method. A transglycosylation reaction of the GlcNAc-modified insulin using Endo-M gave mono-transglycosylated insulin predominantly. We determined the transglycosylation site of the mono-transglycosylated insulin.
Tomabechi
Yusuke
Y
Department of Applied Chemistry, School of Engineering, Tokai University, 1117 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan.
Suzuki
Rena
R
Haneda
Katsuji
K
Inazu
Toshiyuki
T
D000117:Acetylglucosamine / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D000118:Acetylglucosaminidase / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D007328:Insulin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009089:Mucor / Q000201:enzymology*
D007328:Insulin; C039522:insulin, glycosylated; D000118:Acetylglucosaminidase; D000117:Acetylglucosamine