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Enzymic feruloylation of arabinoxylan-trisaccharide by feruloyl-CoA:arabinoxylan-trisaccharide O-hydroxycinnamoyl transferase from Oryza sativa

机译:阿魏酰-CoA:阿拉伯木聚糖-三糖的O-羟基肉桂酰基转移酶对阿拉伯木聚糖-三糖的酶促阿魏酸化

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摘要

Feruloyl-CoA:arabinoxylan-trisaccharide O-hydroxycinnamoyl transferase, which catalyzes the transfer of ferulic acid from Fer-CoA to arabinoxylan-trisaccharide in the formation of feruloyl arabinoxylan-trisaccharide (Fer-AXX), has been found in an ionically bound fraction and a cytosol fraction of suspension-cultured rice (Oriza sativa L. cv. Nipponbare) cells. Analysis of reaction products by high-performance liquid chromatography showed the formation of product A, which is one of the transfer products having the same retention time as authentic Fer-AXX. Product A was purified by reverse-phase chromatographies to characterize its structure. The isolated product A showed the same ultraviolet spectrum and molecular weight on fast atom bombardment mass spectrometric analysis as those of authentic Fer-AXX. Alakaline saponification of product A released ferulic acid and oligosaccharide. The released oligosaccharide consisted of arabinose and xylose in a molar ratio of 1:2. These results support the identity of product A as feruloylated arabinoxylan-trisaccharide and show the existence of a feruloyltransferase catalyzing the feruloylation of a hemicellulosic fragment.
机译:阿魏酰-CoA:阿拉伯木聚糖-三糖的O-羟基肉桂酰基转移酶,可催化阿魏酸从阿魏酸向阿糖基木聚糖-三糖的转移,形成阿魏酸酯-木糖基-三糖(Fer-AXX),并通过离子键结合悬浮培养的水稻(Oriza sativa L. cv。Nipponbare)细胞的胞浆级分。用高效液相色谱法分析反应产物表明形成了产物A,该产物A是与真实的Fer-AXX具有相同保留时间的转移产物之一。通过反相色谱法纯化产物A以表征其结构。在快速原子轰击质谱分析中,分离出的产物A具有与真正的Fer-AXX相同的紫外光谱和分子量。产物A的碱性皂化释放出阿魏酸和低聚糖。释放的寡糖由阿拉伯糖和木糖组成,摩尔比为1:2。这些结果支持了产物A为阿魏酸化的阿拉伯木聚糖-三糖的身份,并表明存在催化半纤维素片段阿魏酸化的阿魏酰转移酶。

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