来自Gloeophyllum trabeum的一种单域膨胀类蛋白质,能够分裂西兰
Ignacio Delgado Santamaría1, Heidi Østby1, Vincent G H Eijsink1
1Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås 1432, Norway.
扩增相关蛋白 (ERP) 曾被认为是非催化性的,但这项研究揭示了真菌ERP中的催化活性. 这一发现挑战了目前对ERP功能在细胞壁修饰中的理解.
科学领域:
- 生物化学 生物化学
- 菌类学 菌类学是指菌类学.
- 植物科学 植物科学
背景情况:
- 扩增相关蛋白 (ERP) 通常被认为是非催化细胞壁放松剂.
- ERP具有类似于GH45内葡萄糖酶的催化域的域,但没有观察到任何催化活性.
- 目前的范式表明,ERP主要通过非催化机制发挥作用.
研究的目的:
- 为了研究来自棕色腐烂真菌Gloeophyllum trabeum的单域ERP的催化活性.
- 为了识别酶产物和描述真菌ERP的催化机制.
- 重新评估对ERP函数的既定理解.
主要方法:
- 使用高性能离子交换染色学与脉冲电压计检测进行酶检测.
- 对烯衍生物产品的分析,包括葡萄糖化烯酸糖.
- 基于结构的ERP和GH45内葡萄糖酶的多重序列对齐.
- 保存的酸性残留物 (Asp87和Asp25) 的位点定向突变发生.
主要成果:
- 由真菌ERP,GtEXPN_133317.证明了由酶依赖的西兰衍生物产品的形成.
- 鉴定出葡萄糖化西-寡糖化物作为酶活性的特定产物.
- 保存的酸性残留物Asp87和Asp25的突变性取消了解活性.
- 菌ERP具有第二种保存的酸性残留物,这种残留物在植物ERP中不存在.
结论:
- 与扩散相关的蛋白质,特别是像GtEXPN_133317这样的真菌ERP,表现出解活性.
- 催化活性依赖于保存的酸性残留物,这表明一种新的酶机制.
- 这一发现挑战了长期以来对ERP的观点,认为ERP是完全非催化性的,并扩大了其功能范围.
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