Xylan-Degrading Catalytic Flagellar Nanorods
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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.
KeywordsFlagellin Xylanase A Polymerization Self-assembly Flagellar export Nanorod
Xylanase A from Bacillus subtilis
Fusion construct of FliC and XynA with a C-terminal His6 tag
Sodium dodecyl sulfate–polyacrylamide gel electrophoresis
Phosphate buffered saline
Flagellin-enzyme fusion protein
We thank K. Namba and P. Závodszky for support and encouragement. We are grateful to the Japan Science and Technology Corporation for generous donation of equipment. This work was supported by the Hungarian Scientific Research Fund (OTKA) grants K104726 and NK108642.
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