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Identification of an arabinopyranosyltransferase from Physcomitrella patens involved in the synthesis of the hemicellulose xyloglucan

机译:鉴定了涉及半纤维素木葡聚糖合成的小立碗藓中的阿拉伯吡喃糖基转移酶

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The hemicellulose xyloglucan consists of a backbone of a β‐1,4 glucan substituted with xylosyl moieties and many other, diverse side chains that are important for its proper function. Many, but not all glycosyltransferases involved in the biosynthesis of xyloglucan have been identified. Here, we report the identification of an hitherto elusive xyloglucan:arabinopyranosyltransferase. This glycosyltransferase was isolated from the moss Physcomitrella patens , where it acts as a xyloglucan “D”‐side chain transferase (XDT). Heterologous expression of PpXDT in the Arabidopsis thaliana double mutant mur3.1 xlt2 , where xyloglucan consists of a xylosylated glucan without further glycosyl substituents, results in the production of the arabinopyranose‐containing “D” side chain as characterized by oligosaccharide mass profiling, glycosidic linkage analysis, and NMR analysis. In addition, expression of a related Physcomitrella glycosyltransferase ortholog of PpXLT2 leads to the production of the galactose‐containing “L” side chain. The presence of the “D” and “L” xyloglucan side chains in the Arabidopsis double mutant Atmur3.1 xlt2 expressing PpXDT and PpXLT2 , respectively, rescues the dwarfed phenotype of untransformed Atmur3.1 xlt2 mutants to nearly wild‐type height. Expression of PpXDT and PpXLT2 in the Atmur3.1 xlt2 mutant also enhanced root growth.
机译:半纤维素木葡聚糖由被木糖基部分取代的β-1,4葡聚糖的骨架和许多其他多样的侧链组成,这些侧链对于其正常功能很重要。已经鉴定出参与木葡聚糖生物合成的许多但不是全部糖基转移酶。在这里,我们报告迄今难以捉摸的木葡聚糖:阿拉伯吡喃糖基转移酶的鉴定。这种糖基转移酶是从苔藓小立碗藓(Physcomitrella patens)中分离出来的,在这里它充当木葡聚糖“ D”侧链转移酶(XDT)。 PpXDT在拟南芥双突变体mur3.1 xlt2中的异源表达,其中木葡聚糖由没有进一步糖基取代基的木糖基化葡聚糖组成,导致产生含阿拉伯吡喃糖的“ D”侧链,其特征为寡糖质量谱分析,糖苷键连接分析和NMR分析。此外,PpXLT2相关的Physcomitrella糖基转移酶直向同源物的表达会导致产生含半乳糖的“ L”侧链。分别表达PpXDT和PpXLT2的拟南芥双突变体Atmur3.1 xlt2中“ D”和“ L”木葡聚糖侧链的存在将未转化的Atmur3.1 xlt2突变体的矮化表型挽救到接近野生型的高度。在Atmur3.1 xlt2突变体中PpXDT和PpXLT2的表达也增强了根的生长。

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