An esterase from the basidiomycete Pleurotus sapidus hydrolyzes feruloylated saccharides
An esterase from the basidiomycete Pleurotus sapidus hydrolyzes feruloylated saccharides
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DOI：
10.1007/s00253-012-4598-7
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发表时间：
2013-08-01
影响因子：
5
通讯作者：
Berger, Ralf G.
中科院分区：
文献类型：
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作者：
Linke, Diana;Matthes, Rene;Berger, Ralf G.
研究担子菌真菌 Pleurotus sapidus 在富含 Tween 80 的营养培养基中分泌的酯酶，发现一种酶可以水解阿魏酰化糖的酯键。通过离子交换和尺寸排阻色谱法纯化酶。聚丙烯酰胺凝胶电泳分析显示约55 kDa的单体蛋白。完整的编码序列具有1,665 bp的开放阅读框，编码由554个氨基酸组成的蛋白质（Est1）。该酶与任何已发表的阿魏酸酯酶序列没有显着同源性，但具有脂肪酶/酯酶超家族的推定保守结构域。底物特异性研究将新酶归类为 A 型阿魏酸酯酶，可水解阿魏酸甲酯、芥子酸甲酯和对香豆酸甲酯，但不水解咖啡酸甲酯。该酶的最适pH值为6，最适温度为50°C。阿魏酸从阿魏糖中有效释放，阿魏酸酯酶在双相系统（50%甲苯或叔丁基甲基醚）中表现出中等稳定性。
Investigating the secretion of esterases by the basidiomycetous fungus Pleurotus sapidus in a Tween 80-rich nutrient medium, an enzyme was discovered that hydrolyzed the ester bond of feruloylated saccharides. The enzyme was purified by ion exchange and size exclusion chromatography. Polyacrylamide gel electrophoresis analysis showed a monomeric protein of about 55 kDa. The complete coding sequence with an open reading frame of 1,665 bp encoded a protein (Est1) consisting of 554 amino acids. The enzyme showed no significant homology to any published feruloyl esterase sequences, but possessed putative conserved domains of the lipase/esterase superfamily. Substrate specificity studies classified the new enzyme as type-A feruloyl esterase, hydrolyzing methyl ferulate, methyl sinapate, and methyl p-coumarate but no methyl caffeate. The enzyme had a pH optimum of 6 and a temperature optimum at 50 A degrees C. Ferulic acid was efficiently released from ferulated saccharides, and the feruloyl esterase exhibited moderate stability in biphasic systems (50 % toluene or tert-butylmethyl ether).