来自 Dichostereum sordulentum 1488 的 α-岩藻糖苷酶的纯化和表征。

IF 2.4 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Carbohydrate Research Pub Date : 2024-09-17 DOI:10.1016/j.carres.2024.109278

Lorena Herrera , María Eugenia Cedrés , Paula Rodríguez Bonnecarrere , Cecilia Giacomini

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引用次数: 0

摘要

生物糖介导多种生理过程，因此糖基化模式的改变可导致不同的疾病，如自身免疫性疾病、传染性疾病、慢性抗炎性疾病，甚至癌症。事实上，N-或O-聚糖中的岩藻糖基化改变是与癌症相关的糖基化模式中最常见的变化之一。因此，阐明糖共轭聚糖的作用对于了解涉及糖共轭聚糖的病理发展至关重要。从这个意义上说，糖苷酶是极好的工具，因为它们能催化糖残基的选择性去除，从而可以评估糖基去除导致的生物作用变化。这项研究描述了从真菌 Dichostereum sordulentum 1488 中纯化出的α-岩藻糖苷酶的特性。它是一种同源二聚体，分子量为 214 kDa，最适 pH 值和温度分别为 4.0 和 70 °C。它对底物对硝基苯-α-l-岩藻糖苷的 KM 值为 0.27 mM，VMax 值为 3.3 μmoles PNP/min/mg，显示出底物抑制特征。它对水解以 α-(1,2)键连接的岩藻糖具有高度特异性。这种新的α-岩藻糖苷酶的鉴定、纯化和表征对于扩大糖苷酶作为聚糖阐释工具的可用性具有重要意义。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Purification and characterization of α-fucosidase from Dichostereum sordulentum 1488

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Purification and characterization of α-fucosidase from Dichostereum sordulentum 1488

Biological glycans mediate several physiological processes, thus altered glycosylation patterns can lead to different diseases such as autoimmune, infectious, chronic anti-inflammatory diseases, or even cancer. In fact, alterations in fucosylation in either N- or O-glycans are among the most frequent changes in glycosylation patterns associated with cancer. Therefore, elucidation of the role of glycoconjugate glycans is essential for understanding the development of pathologies where they are involved. In this sense glycosidases are excellent tools, since they catalyse the selective removal of sugar residues, allowing the evaluation of changes in their biological role due to glycan removal. This work describes the purification and characterization of a α-fucosidase from the fungus Dichostereum sordulentum 1488. It is a homodimer with a molecular weight of 214 kDa and optimum pH and temperature of 4.0 and 70 °C respectively. It has a K_M of 0.27 mM and V_Max of 3.3 μmoles PNP/min per mg for the substrate p-nitrophenyl-α-l-fucopyranoside, showing a substrate inhibition profile. It showed high specificity for the hydrolysis of fucose linked by α-(1,2) bonds. The identification, purification, and characterization of this new α-fucosidase is highly relevant for enlarging the availability of glycosidases for use as tools for glycan elucidation.

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来源期刊

Carbohydrate Research 化学-生化与分子生物学

CiteScore

5.00

自引率

3.20%

发文量

183

审稿时长

3.6 weeks

期刊介绍： Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects. Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence. Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".