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6B血清型WciN及其突变体的生化特性与合成应用

Biochemical Characterization and Synthetic Application of WciN and Its Mutants From Serotype 6B.

作者信息

Gong Wei, Liang Min, Zhao Jielin, Wang Hong, Chen Zonggang, Wang Fengshan, Gu Guofeng

机构信息

National Glycoengineering Research Center, Shandong Provincial Key Laboratory of Carbohydrate Chemistry and Glycobiology, Shandong University, Qingdao, China.

School of Pharmaceutical Science, Shandong University, Jinan, China.

出版信息

Front Chem. 2022 Jun 15;10:914698. doi: 10.3389/fchem.2022.914698. eCollection 2022.

DOI:10.3389/fchem.2022.914698
PMID:35783203
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9240355/
Abstract

The biochemical properties of α-1,3-galactosyltransferase WciN from serotype 6B were systemically characterized with the chemically synthesized Glcα-PP-(CH)-OPh as an acceptor substrate. The site-directed mutation of D38 and A150 residues of WciN was further investigated, and the enzymatic activities of those WciN mutants revealed that A150 residue was the pivotal residue responsible for nucleotide donor recognition and the single-site mutation could completely cause pneumococcus serotype switch. Using WciN and WciN mutants as useful tool enzymes, the disaccharides Galα1,3Glcα-PP-(CH)-OPh and Glcα1,3Glcα-PP-(CH)-OPh were successfully prepared in multi-milligram scale in high yields.

摘要

以化学合成的Glcα-PP-(CH)-OPh作为受体底物，系统地表征了6B血清型α-1,3-半乳糖基转移酶WciN的生化特性。进一步研究了WciN的D38和A150残基的定点突变，这些WciN突变体的酶活性表明，A150残基是负责核苷酸供体识别的关键残基，单点突变可完全导致肺炎球菌血清型转换。使用WciN和WciN突变体作为有用的工具酶，成功地以高产率制备了多毫克规模的二糖Galα1,3Glcα-PP-(CH)-OPh和Glcα1,3Glcα-PP-(CH)-OPh。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/1d1a101a78f1/fchem-10-914698-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/931173a3d685/fchem-10-914698-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/2ddbbe97376b/fchem-10-914698-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/718fbee2ea3e/fchem-10-914698-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/d0bb871373c2/fchem-10-914698-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/787b8290dd50/fchem-10-914698-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/1d1a101a78f1/fchem-10-914698-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/931173a3d685/fchem-10-914698-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/2ddbbe97376b/fchem-10-914698-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/718fbee2ea3e/fchem-10-914698-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/d0bb871373c2/fchem-10-914698-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/787b8290dd50/fchem-10-914698-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3559/9240355/1d1a101a78f1/fchem-10-914698-g006.jpg

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本文引用的文献

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New Pneumococcal Serotype 15D.新型肺炎球菌血清型15D
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A New Pneumococcal Capsule Type, 10D, is the 100th Serotype and Has a Large Fragment from an Oral Streptococcus.
一种新型肺炎球菌荚膜类型 10D 是第 100 种血清型，具有来自口腔链球菌的大片段。
mBio. 2020 May 19;11(3):e00937-20. doi: 10.1128/mBio.00937-20.
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Correlates of Nonrandom Patterns of Serotype Switching in Pneumococcus.肺炎球菌血清型转换非随机模式的相关因素。
J Infect Dis. 2020 Apr 27;221(10):1669-1676. doi: 10.1093/infdis/jiz687.
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Biochemical studies of a β-1,4-rhamnoslytransferase from Streptococcus pneumonia serotype 23F.肺炎链球菌 23F 型β-1,4-鼠李糖基转移酶的生化研究。
Org Biomol Chem. 2019 Jan 31;17(5):1071-1075. doi: 10.1039/c8ob02795a.
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Pneumococcal Capsule Synthesis Locus cps as Evolutionary Hotspot with Potential to Generate Novel Serotypes by Recombination.肺炎球菌荚膜合成基因座cps作为具有通过重组产生新型血清型潜力的进化热点。
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