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Characterization and application of active human α2,6-sialyltransferases ST6GalNAc V and ST6GalNAc VI recombined in Escherichia coli

文献类型：期刊论文

作者Pei, Caixia4,5,6; Peng, Xinlv2,4,5; Wu, Yiran2,4,5; Jiao, Runmiao4,5,6; Li, Tiehai3; Jiao, Siming1,4,5; Zhou, Lei1,4,5; Li, Jianjun1,4,5; Du, Yuguang1,4,5; Qian, Eika W.6
刊名ENZYME AND MICROBIAL TECHNOLOGY
出版日期2024-06-01
卷号177页码:10
关键词N-acetylgalactosaminide alpha 2,6-sialyltransferase hST6GalNAc V hST6GalNAc VI Disialyllacto-N-tetraose (DSLNT) Chaperone Escherichia coli
ISSN号0141-0229
DOI10.1016/j.enzmictec.2024.110426
通讯作者Li, Jianjun(jjli@ipe.ac.cn) ; Du, Yuguang(ygdu@ipe.ac.cn) ; Qian, Eika W.(whqian@cc.tuat.ac.jp)
英文摘要Eukaryotic sialyltransferases play key roles in many physiological and pathological events. The expression of active human recombinant sialyltransferases in bacteria is still challenging. In the current study, the genes encoding human N-acetylgalactosaminide alpha 2,6-sialyltransferase V (hST6GalNAc V) and N-acetylgalactosaminide alpha 2,6-sialyltransferase VI (hST6GalNAc VI) lacking the N-terminal transmembrane domains were cloned into the expression vectors, pET-32a and pET-22b, respectively. Soluble and active forms of recombinant hST6GalNAc V and hST6GalNAc VI when coexpressed with the chaperone plasmid pGro7 were successfully achieved in Escherichia coli. Further, lactose (Lac), Lacto-N-triose II (LNT II), lacto-N-tetraose (LNT), and sialyllacto-N-tetraose a (LSTa) were used as acceptor substrates to investigate their activities and substrate specificities. Unexpectedly, both can transfer sialic acid onto all those substrates. Compared with hST6GalNAc V expressed in the mammalian cells, the recombinant two alpha 2,6-sialyltransferases in bacteria displayed flexible substrate specificities and lower enzymatic efficiency. In addition, an important human milk oligosaccharide disialyllacto-N-tetraose (DSLNT) can be synthesized by both human alpha 2,6-sialyltransferases expressed in E. coli using LSTa as an acceptor substrate. To the best of our knowledge, these two active human alpha 2,6-sialyltransferases enzymes were expressed in bacteria for the first time. They showed a high potential to be applied in biotechnology and investigating the molecular mechanisms of biological and pathological interactions related to sialylated glycoconjugates.
WOS关键词NECROTIZING ENTEROCOLITIS ; SIALIC ACIDS ; SIALYLTRANSFERASE ; EXPRESSION ; GLYCOSYLATION ; REVEALS
资助项目Foundation of the Natural Science Foundation of China[21877114] ; Innovation Academy for Green Manufacture Institute, Chinese Academy of Sciences[IAGM2020C31]
WOS研究方向Biotechnology & Applied Microbiology
语种英语
WOS记录号WOS:001218643800001
出版者ELSEVIER SCIENCE INC
源URL[http://119.78.100.183/handle/2S10ELR8/311279]  
专题新药研究国家重点实验室
通讯作者Li, Jianjun; Du, Yuguang; Qian, Eika W.
作者单位1.Chinese Acad Sci, Innovat Acad Green Manufacture Inst, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Sch Chem Engn, Beijing 100049, Peoples R China
3.Chinese Acad Sci, Shanghai Inst Mat Med, Carbohydrate Based Drug Res Ctr, CAS Key Lab Receptor Res,State Key Lab Drug Res, Shanghai 201203, Peoples R China
4.Chinese Acad Sci, Key Lab Biopharmaceut Preparat & Delivery, Beijing 100190, Peoples R China
5.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China
6.Tokyo Univ Agr & Technol, Grad Sch Bioapplicat & Syst Engn, Nakacho 2-24-16, Tokyo 1848588, Japan
推荐引用方式
GB/T 7714		 Pei, Caixia,Peng, Xinlv,Wu, Yiran,et al. Characterization and application of active human α2,6-sialyltransferases ST6GalNAc V and ST6GalNAc VI recombined in Escherichia coli[J]. ENZYME AND MICROBIAL TECHNOLOGY,2024,177:10.
APA Pei, Caixia.,Peng, Xinlv.,Wu, Yiran.,Jiao, Runmiao.,Li, Tiehai.,...&Qian, Eika W..(2024).Characterization and application of active human α2,6-sialyltransferases ST6GalNAc V and ST6GalNAc VI recombined in Escherichia coli.ENZYME AND MICROBIAL TECHNOLOGY,177,10.
MLA Pei, Caixia,et al."Characterization and application of active human α2,6-sialyltransferases ST6GalNAc V and ST6GalNAc VI recombined in Escherichia coli".ENZYME AND MICROBIAL TECHNOLOGY 177(2024):10.

入库方式： OAI收割

来源：上海药物研究所

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