Department of Applied Chemistry, School of Engineering, Tokai University, 1117 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan.
Bioorg Med Chem. 2010 Feb;18(3):1259-64. doi: 10.1016/j.bmc.2009.12.031. Epub 2009 Dec 16.
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.
采用酶化学法，利用毛霉（Mucor hiemalis）内切-β-N-乙酰氨基葡萄糖苷酶（Endo-M）合成具有 N-连接寡糖的人工胰岛素。采用 Mpt-MA 法，通过 GlcNAc 羧甲基糖苷与牛胰岛素 3 个氨基的反应，制备 GlcNAc 修饰的胰岛素。用 Endo-M 对 GlcNAc 修饰的胰岛素进行转糖基化反应，主要得到单转糖基化胰岛素。我们确定了单转糖基化胰岛素的转糖基化位点。
