Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616, USA.
Bioorg Med Chem. 2013 Aug 15;21(16):4778-85. doi: 10.1016/j.bmc.2013.02.040. Epub 2013 Mar 7.
A series of STn-MUC1 and ST-MUC1 glycopeptides containing naturally occurring and non-natural sialic acids have been chemoenzymatically synthesized from Tn-MUC1 glycopeptide using one-pot multienzyme (OPME) approaches. In situ generation of the sialyltransferase donor cytidine 5'-monophosphate-sialic acid (CMP-Sia) using a CMP-sialic acid synthetase in the presence of an extra amount of cytidine 5'-triphosphate (CTP) and removal of CMP from the reaction mixture by flash C18 cartridge purification allow the complete consumption of Tn-MUC1 glycopeptide for quantitative synthesis of STn-MUC1. A Campylobacter jejuni β1-3GalT (CjCgtBΔ30-His6) mutant has been found to catalyze the transfer of one or more galactose residues to Tn-MUC1 for the synthesis of T-MUC1 and galactosylated T-MUC1. Sialylation of T-MUC1 using Pasteurella multocida α2-3-sialyltransferase 3 (PmST3) with Neisseria meningitidis CMP-sialic acid synthetase (NmCSS) and Escherichia coli sialic acid aldolase in one pot produced ST-MUC1 efficiently. These glycopeptides are potential cancer vaccine candidates.
已通过一锅多酶（OPME）方法，从 Tn-MUC1 糖肽化学酶促合成了一系列含有天然和非天然唾液酸的 STn-MUC1 和 ST-MUC1 糖肽。在存在额外量的胞苷 5′-三磷酸（CTP）的情况下，使用 CMP-唾液酸合成酶原位生成唾液酸转移酶供体胞苷 5′-单磷酸唾液酸（CMP-Sia），并通过快速 C18 柱纯化从反应混合物中除去 CMP，从而允许 Tn-MUC1 糖肽完全消耗，用于 STn-MUC1 的定量合成。已经发现弯曲杆菌属空肠β1-3GalT（CjCgtBΔ30-His6）突变体能够催化一个或多个半乳糖残基向 Tn-MUC1 的转移，用于合成 T-MUC1 和半乳糖化的 T-MUC1。使用多杀巴斯德氏菌α2-3-唾液酸转移酶 3（PmST3）与脑膜炎奈瑟氏球菌 CMP-唾液酸合成酶（NmCSS）和大肠杆菌唾液酸醛缩酶在一锅法中对 T-MUC1 进行唾液酸化，有效地产生了 ST-MUC1。这些糖肽是潜在的癌症疫苗候选物。
