Protein nanofibrils self-assembled from crystallin proteins, a waste material from the fishing industry, have been identified as a suitable material to be used as a bionanoscaffold. In this study, a successful method for the functionalization of crystallin protein nanofibrils by immobilizing several enzymes of industrial relevance (glucose oxidase, β-galactosidase, pectinase, α-amylase, and laccase) through a glutaraldehyde cross-linking approach is reported. The extent of functionalization is evaluated by using gel electrophoresis, transmission electron microscopy, and thermostability studies. The functionalized fibrils are investigated further for reusability studies-the use of protein nanofibrils as nanoscaffolds results in a significant increase in enzyme thermostability and reusability relative to the free enzyme in solution under the same conditions. Finally, as an example and proof of concept, the use of the developed functionalization method in a biosensor platform for glucose and lactose detection is shown, by utilizing the crystallin protein nanofibrils as nanoscaffolds.
Keywords: biosensors; enzymes; fibrous proteins; immobilization; nanostructures.
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