NMR-Based Structural Glycomics for High-Throughput Screening of Carbohydrate-Active Enzyme Specificity

被引：27

作者：

Irague, Romain ^{[1
,2
,3
]}

Massou, Stephane ^{[1
,2
,3
]}

Moulis, Claire ^{[1
,2
,3
]}

Saurel, Olivier ^{[4
]}

Milon, Alain ^{[4
]}

Monsan, Pierre ^{[1
,2
,3
]}

Remaud-Simeon, Magali ^{[1
,2
,3
]}

Portais, Jean-Charles ^{[1
,2
,3
]}

Potocki-Veronese, Gabrielle ^{[1
,2
,3
]}

机构：

[1] Univ Toulouse, INSA, UPS, INP,LISBP, F-31077 Toulouse, France

[2] INRA, CNRS, UMR5504, F-31400 Toulouse, France

[3] INRA, CNRS, Ingn Syst Biol & Proc UMR792, F-31400 Toulouse, France

[4] Inst Pharmacol & Biol Struct, CNRS, UPS, Toulouse, France

来源：

ANALYTICAL CHEMISTRY | 2011年 / 83卷 / 04期

关键词：

ALPHA-D-GLUCANS; CONSTRUCTION; SPECTROSCOPY; LIBRARIES; TOOL;

D O I：

10.1021/ac1032148

中图分类号：

O65 [分析化学];

学科分类号：

070302 [分析化学];

摘要：

We report here the development of a straightforward, sensitive, and quantitative NMR-based method for high-throughput characterization of carbohydrate structure and screening of carbohydrate active enzyme (CAZyme) specificity. Automated assays starting from gene library expression to carbohydrate structure determination directly from crude reaction media have been established and successfully used to screen a library of 4032 CAZymes obtained by combinatorial engineering, at a rate of 480 enzyme variants per day. This allowed one to accurately discriminate 303 enzyme variants with altered specificity. The results demonstrate the potential of high-throughput NMR technology in novel biocatalysts in glycomics, to mine artificial and natural enzyme diversity for

引用

页码：1202 / 1206

页数：5

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