Successfully Engineering a Bacterial Sialyltransferase for Regioselective alpha 2,6-sialylation
Successfully Engineering a Bacterial Sialyltransferase for Regioselective alpha 2,6-sialylation
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成功设计用于区域选择性 α 2,6-唾液酸化的细菌唾液酸转移酶
DOI：
10.1021/acscatal.8b01993
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发表时间：
2018
期刊：
影响因子：
12.9
通讯作者：
Cheng Jiansong
中科院分区：
文献类型：
--
作者：
Xu Yangyang;Fan Yueyuan;Ye Jinfeng;Wang Faxing;Nie Qu;eng;Wang Li;Wang Peng George;Cao Hongzhi;Cheng Jiansong
本文对丹色光杆菌β-半乳糖苷α2，6-唾液酸转移酶(Pd2，6ST)进行了重新设计，该酶能同时分解末端和内部半乳糖和N-乙酰半乳糖胺，用于区域选择性地产生末端α2，6-唾液酸苷。在最近开发的凹坑策略的指导下，在Ala200和Ser232位点产生了一系列突变，以重塑受体结合口袋。Pd2，6ST双突变体A200Y/S232Y的受体结合口袋为L状，是一种优势的α2，6-唾液酸基转移酶，能有效催化半乳糖或乙酰半乳糖胺在一系列糖链的非还原末端的区域选择性α-2，6-唾液酸化反应。同时，A200Y/S232Y保持了灵活的供体底物特异性，能够转移Neu5Ac、Neu5Gc和KdN。
A β-galactoside α2,6-sialyltransferase fromPhotobacterium damselae(Pd2,6ST) that is capable of sialylating both terminal and internal galactose andN-acetylgalactosamine was herein redesigned for regioselectively producing terminal α2,6-sialosides. Guided by a recently developed bump-hole strategy, a series of mutations at Ala200 and Ser232 sites were created for reshaping the acceptor binding pocket. Finally, a Pd2,6ST double mutant A200Y/S232Y with an altered L-shaped acceptor binding pocket was identified to be a superior α2,6-sialyltransferase which can efficiently catalyze the regioselective α2,6-sialylation of galactose orN-acetylgalactosamine at the nonreducing end of a series of glycans. Meanwhile, A200Y/S232Y remains flexible donor substrate specificity and is able to transfer Neu5Ac, Neu5Gc, and KDN.