Klein Ágnes, Szabó Veronika, Kovács Mátyás, Patkó Dániel, Tóth Balázs, Vonderviszt Ferenc
Bio-Nanosystems Laboratory, Research Institute of Chemical and Process Engineering, Faculty of Information Technology, University of Pannonia, Egyetem u. 10, Veszprém, 8200, Hungary.
Mol Biotechnol. 2015 Sep;57(9):814-9. doi: 10.1007/s12033-015-9874-1.
Flagellin, the main component of flagellar filaments, is a protein possessing polymerization ability. In this work, a novel fusion construct of xylanase A from B. subtilis and Salmonella flagellin was created which is applicable to build xylan-degrading catalytic nanorods of high stability. The FliC-XynA chimera when overexpressed in a flagellin deficient Salmonella host strain was secreted into the culture medium by the flagellum-specific export machinery allowing easy purification. Filamentous assemblies displaying high surface density of catalytic sites were produced by ammonium sulfate-induced polymerization. FliC-XynA nanorods were resistant to proteolytic degradation and preserved their enzymatic activity for a long period of time. Furnishing enzymes with self-assembling ability to build catalytic nanorods offers a promising alternative approach to enzyme immobilization onto nanostructured synthetic scaffolds.
鞭毛蛋白是鞭毛丝的主要成分，是一种具有聚合能力的蛋白质。在这项工作中，构建了一种来自枯草芽孢杆菌的木聚糖酶A与沙门氏菌鞭毛蛋白的新型融合体，可用于构建高稳定性的木聚糖降解催化纳米棒。当FliC-XynA嵌合体在鞭毛蛋白缺陷的沙门氏菌宿主菌株中过表达时，它通过鞭毛特异性输出机制分泌到培养基中，便于纯化。通过硫酸铵诱导聚合产生了具有高催化位点表面密度的丝状聚集体。FliC-XynA纳米棒对蛋白水解降解具有抗性，并能长时间保持其酶活性。赋予酶自组装能力以构建催化纳米棒为将酶固定在纳米结构合成支架上提供了一种有前景的替代方法。
