One-pot multienzyme (OPME) chemoenzymatic synthesis of brain ganglioside glycans with human ST3GAL II expressed in E. coli.
One-pot multienzyme (OPME) chemoenzymatic synthesis of brain ganglioside glycans with human ST3GAL II expressed in E. coli.
复制标题
使用大肠杆菌中表达的人 ST3GAL II 进行脑神经节苷脂聚糖的一锅多酶 (OPME) 化学酶合成。
DOI：
10.1002/cctc.202101498
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发表时间：
2022
期刊：
影响因子：
4.5
通讯作者：
Chen,Xi
中科院分区：
文献类型：
--
作者：
Yang,Xiaoxiao;Yu,Hai;Yang,Xiaohong;Kooner,AnoopjitSingh;Yuan,Yue;Luu,Bryant;Chen,Xi
人唾液酸转移酶ST 3GAL II（hST 3GAL II）在大肠杆菌中成功表达，得到了一种具有活性的可溶性融合蛋白，其N端为麦芽糖结合蛋白（MBP），C端为六组氨酸标签。 它被用作一锅多酶（OPME）唾液酸化系统中的有效催化剂，用于高产量生产神经节苷脂GM 1b和高唾液酸化脑神经节苷脂GD 1a和GT 1b的聚糖。在另一个OPME唾液酸化反应中使用细菌α2-8-唾液酸转移酶进一步唾液酸化GM 1b和GD 1a聚糖，分别形成GD 1c和脑神经节苷脂GT 1a聚糖。与其细菌唾液酸转移酶对应物相比，重组hST 3GAL II的较低的反向糖基化活性简化了酶促合成反应的处理，并且对于将来在自动化化学酶促合成过程中的使用具有优势。
A human sialyltransferase ST3GAL II (hST3GAL II) was successfully expressed inEscherichia colias an active soluble fusion protein with an N‐terminal maltose‐binding protein (MBP) and a C‐terminal hexa‐histidine tag. It was used as an efficient catalyst in a one‐pot multienzyme (OPME) sialylation system for high‐yield production of the glycans of ganglioside GM1b and highly sialylated brain gangliosides GD1a and GT1b. Further sialylation of GM1b and GD1a glycans using a bacterial α2–8‐sialyltransferase in another OPME sialylation reaction led to the formation of the glycans of GD1c and brain ganglioside GT1a, respectively. The lower reverse glycosylation activity of the recombinant hST3GAL II compared to its bacterial sialyltransferase counterpart simplifies the handling of enzymatic synthetic reactions and has an advantage for future use in automated chemoenzymatic synthetic processes.