山东大学曹鸿志团队开发了用于位点特异性酶聚焦化的氧化还原控制底物工程策略。相关研究成果于2022年10月17日发表在《德国应用化学》。

岩藻糖基化是寡聚N-乙酰基乳糖胺（寡聚LacNAc）聚糖最常见的修饰之一。然而，没有一种已知的岩藻糖基转移酶（FucTs）能够以位点特异的方式，将a1,3-连接岩藻糖安装到寡聚LacNAc底物上。

该文中，研究人员报告了一种简单且通用的氧化还原控制底物工程策略，用于含有多个LacNAc单元的复杂寡糖的位点特异性a1,3-糖基化。该策略利用操作简单的氧化酶模块，通过使用半乳糖氧化酶（GOase）将LacNAc单元转化为氧化的C6’-醛LacNAc序列。与LacNAc相比，氧化的C6’-醛LacNAc序列不是幽门螺杆菌重组a1,3-葡糖基转移酶（Hpa1,3FucT）的良好底物，因此能够在完整的LacNAd位点实现位点特异性a1,3-糖基化。通过合成多种结构明确的直链和支链O-和N-连接聚糖的岩藻糖苷，证明了这种氧化还原控制底物工程策略的普适性和鲁棒性。

附：英文原文

Title: A Redox-Controlled Substrate Engineering Strategy for Site-Specific Enzymatic Fucosylation

Author: Na Lu, Yu Li, Hui Xia, Kan Zhong, Chenning Jia, Jinfeng Ye, Xianwei Liu, Chang-Cheng Liu, Hongzhi Cao

Issue&Volume: 2022-10-17

Abstract: Fucosylation is one of the most common modifications of oligo-N-acetyllactosamine (oligo-LacNAc) glycans. However, none of the known fucosyltransferases (FucTs) could install the a1,3-linked fucose to the oligo-LacNAc substrates in a site-specific manner. Here, we report a facile and general redox-controlled substrate engineering strategy for the site-specific a1,3-fucosylation of complex oligosaccharides containing multiple LacNAc units. This strategy takes advantage of an operationally simple oxidation enzyme module by using galactose oxidase (GOase) to convert the LacNAc unit into oxidized C6'-aldehyde LacNAc sequence. In contrast with LacNAc, the oxidized C6'-aldehyde LacNAc sequence is not a good substrate for recombinant a1,3-fucosyltransferase from Helicobacter pylori (Hpa1,3FucT), enabling the site-specific a1,3-fucosylation at intact LacNAc sites. The general applicability and robustness of this redox-controlled substrate engineering strategy was demonstrated by the synthesis of a variety of structurally well-defined fucosides of linear and branched O- and N-linked glycans

DOI: 10.1002/anie.202211032

Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202211032