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快速定量活性分析表明,霍乱弧菌定植因子 GbpA 是一种活性裂解多糖单加氧酶。

A rapid quantitative activity assay shows that the Vibrio cholerae colonization factor GbpA is an active lytic polysaccharide monooxygenase.

机构信息

Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, N-1432 Aas, Norway.

Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, Groningen, The Netherlands.

出版信息

FEBS Lett. 2014 Sep 17;588(18):3435-40. doi: 10.1016/j.febslet.2014.07.036. Epub 2014 Aug 7.

Abstract

The discovery of the copper-dependent lytic polysaccharide monooxygenases (LPMOs) has revealed new territory for chemical and biochemical analysis. These unique mononuclear copper enzymes are abundant, suggesting functional diversity beyond their established roles in the depolymerization of biomass polysaccharides. At the same time basic biochemical methods for characterizing LPMOs, such as activity assays are not well developed. Here we describe a method for quantification of C1-oxidized chitooligosaccharides (aldonic acids), and hence LPMO activity. The method was used to quantify the activity of a four-domain LPMO from Vibriocholerae, GbpA, which is a virulence factor with no obvious role in biomass processing.

摘要

铜依赖型溶菌多糖单加氧酶(LPMOs)的发现揭示了化学和生化分析的新领域。这些独特的单核铜酶丰富多样,表明其功能不仅限于在生物质多糖的解聚中发挥作用。与此同时,用于表征 LPMOs 的基本生化方法,如活性测定,尚未得到很好的发展。在这里,我们描述了一种定量 C1-氧化壳寡糖(醛酸)的方法,从而定量 LPMO 的活性。该方法用于定量霍乱弧菌的四结构域 LPMO,GbpA,其是一种毒力因子,在生物质处理中没有明显作用。

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