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筛选旱金莲(旱金莲属)提取物中的异源转糖基化活性。

Screening for hetero-transglycosylating activities in extracts from nasturtium (Tropaeolum majus).

作者信息

Mohand Fairouz Ait, Farkas Vladimír

机构信息

Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, 84538 Bratislava, Slovakia.

出版信息

Carbohydr Res. 2006 Apr 10;341(5):577-81. doi: 10.1016/j.carres.2006.01.018. Epub 2006 Jan 30.

Abstract

Using combinations of different polysaccharides as glycosyl donors and of oligosaccharides fluorescently labeled by sulforhodamine (SR) as glycosyl acceptors, we screened for the presence of transglycosylating activities in extracts from nasturtium (Tropaeolum majus). Besides xyloglucan endotransglycosylase/hydrolase (XTH/XET, EC 2.4.1.207) activity, which transfers xyloglucanosyl residues from xyloglucan (XG) to XG-derived oligosaccharides (XGOs), a glycosyl transfer from XG to SR-labeled cellooligosaccharides and laminarioligosaccharides has been detected. The XGOs also served as acceptors for the glycosyl transfer from soluble cellulose derivatives carboxymethyl cellulose and hydroxyethylcellulose. The effectivity of these polysaccharides as glycosyl donors for transfer to XG-derived octasaccharide [1-3H]XXLGol decreased in the order XG > HEC > CMC. Isoelectric focusing in polyacrylamide gels showed that bands corresponding to hetero-transglycosylase activities coincided with zones corresponding to XTH/XET. These results can be explained as due either to substrate non-specificity of certain isoenzymes of XTH/XET or to existence of enzymes catalyzing a hetero-transfer, that is the formation of covalent linkages between different types of carbohydrate polymers.

摘要

我们以不同的多糖组合作为糖基供体,以磺基罗丹明(SR)荧光标记的寡糖作为糖基受体,筛选了旱金莲(Tropaeolum majus)提取物中是否存在转糖基活性。除了木葡聚糖内转糖基酶/水解酶(XTH/XET,EC 2.4.1.207)活性(该活性将木葡聚糖残基从木葡聚糖(XG)转移至木葡聚糖衍生的寡糖(XGOs))外,还检测到了从XG到SR标记的纤维寡糖和海带寡糖的糖基转移。XGOs也作为糖基受体,接受来自可溶性纤维素衍生物羧甲基纤维素和羟乙基纤维素的糖基转移。这些多糖作为糖基供体转移至木葡聚糖衍生的八糖[1-³H]XXLGol的有效性顺序为XG>HEC>CMC。在聚丙烯酰胺凝胶中进行等电聚焦显示,与异源转糖基酶活性对应的条带与对应于XTH/XET的区域重合。这些结果可以解释为,要么是由于XTH/XET某些同工酶的底物非特异性,要么是由于催化异源转移(即不同类型碳水化合物聚合物之间形成共价键)的酶的存在。

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