Department of Chemistry, Biotechnology, and Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University, 1-21-40 Korimoto, Kagoshima 890-0065, Japan.
Carbohydr Res. 2010 Mar 30;345(5):631-6. doi: 10.1016/j.carres.2010.01.001. Epub 2010 Jan 7.
This paper describes the phosphorylase-catalyzed enzymatic N-formyl-alpha-glucosaminylation of maltooligosaccharides for direct incorporation of 2-deoxy-2-formamido-alpha-D-glucopyranose units into maltooligosaccharides. When the reaction of 2-deoxy-2-formamido-alpha-D-glucopyranose-1-phosphate (GlcNF-1-P) as the glycosyl donor and maltotetraose as a glycosyl acceptor was performed in the presence of phosphorylase, the N-formyl-alpha-D-glucosaminylated pentasaccharide was produced, as confirmed by MALDI-TOF MS. Furthermore, the glucoamylase-catalyzed reaction of the crude products supported that the 2-deoxy-2-formamido-alpha-D-glucopyranoside unit was positioned at the non-reducing end of the pentasaccharide. The pentasaccharide was isolated from the crude products and its structure was further determined by the (1)H NMR analysis. On the other hand, when the phosphorylase-catalyzed reactions of maltotriose and maltopentaose using GlcNF-1-P were conducted, no N-formyl-alpha-glucosaminylation took place in the former system, whereas the latter system gave N-formyl-alpha-D-glucosaminylated oligosaccharides with various degrees of polymerization. These results could be explained by the recognition behavior of phosphorylase toward maltooligosaccharides.
本文描述了磷酸化酶催化的麦芽寡糖的 N-甲酰基-α-葡糖胺基化反应，用于直接将 2-脱氧-2-甲酰基-α-D-葡吡喃糖基单元掺入麦芽寡糖中。当以 2-脱氧-2-甲酰基-α-D-葡糖胺-1-磷酸（GlcNF-1-P）作为糖基供体，麦芽四糖作为糖基受体进行反应时，在磷酸化酶的存在下，生成了 N-甲酰基-α-D-葡糖胺化五糖，这一点通过 MALDI-TOF MS 得到了证实。此外，粗产物的葡糖淀粉酶催化反应表明，2-脱氧-2-甲酰基-α-D-葡糖苷单元位于五糖的非还原端。从粗产物中分离出五糖，并通过（1）H NMR 分析进一步确定其结构。另一方面，当使用 GlcNF-1-P 进行麦芽三糖和麦芽五糖的磷酸化酶催化反应时，在前一体系中没有发生 N-甲酰基-α-葡糖胺基化，而在后一体系中则得到了具有不同聚合度的 N-甲酰基-α-D-葡糖胺化寡糖。这些结果可以通过磷酸化酶对麦芽寡糖的识别行为来解释。
