Immobilisation of tobacco etch virus (TEV) protease on a high surface area protein nanofibril scaffold. (2018)
AuthorsRaynes JK, Domigan LJ, Pearce FG, Gerrard JAshow all
Tobacco etch virus (TEV) protease is widely used for the removal of poly-histidine affinity tags from proteins. In solution, it is a one-time use enzyme for tag cleavage that has low stability, and is therefore a good candidate for immobilisation. Amyloid fibrils can act as a versatile nanoscaffold by providing a large surface area for biomolecule immobilisation. Immobilisation of TEV protease to amyloid fibrils grown from the surface of a small glass bead, using physisorption, successfully immobilised active TEV protease. The bead retained activity over several uses and successfully cleaved a poly-histidine tag from several his-tagged proteins. This is first time that TEV protease has been immobilised to insulin amyloid fibrils, or any protein based support. Such functionalised surface assembled amyloid fibrils show promise as a novel nanosupport for the creation of functional bionanomaterials, for example, active surface coatings for the production of fine chemicals, chemical detoxification, or biosensing. Insulin amyloid fibrils provide a new nanosupport for the immobilisation of TEV protease, which could allow for the reuse of the enzyme, saving on production costs for recombinantly expressed poly-histidine tagged proteins. This article is protected by copyright. All rights reserved.
ANZSRC Fields of Research06 - Biological Sciences::0601 - Biochemistry and Cell Biology::060107 - Enzymes
06 - Biological Sciences::0601 - Biochemistry and Cell Biology::060109 - Proteomics and Intermolecular Interactions (excl. Medical Proteomics)