title: Vmh2 hydrophobin as a tool for the development of "self-immobilizing" enzymes for biosensing.
abstract: Self-assembling proteins forming amyloid fibrils are promising candidates for the fabrication of biomaterials, due to the chemical and mechanical stability of their structures. Among potential applications, their use as platforms for enzyme immobilization is rapidly gathering attention. In this work, we demonstrate that the production of the enzyme glutathione-S-transferase (GST) fused to the class I hydrophobin Vmh2 from Pleurotus ostreatus represents an invaluable tool for the development of self-immobilizing enzymes useful for high throughput analyses. The proposed immobilization strategy is versatile since it can be applied, in principle, to every recombinant protein able to refold from Escherichia coli inclusion bodies. A GST based biosensor has been developed to quantify toxic compounds, such as the pesticides molinate and captan, in aqueous environmental samples. The main advantages of this sensor include simplicity and speed of preparation, high sensitivity, reusability, and accuracy. Biotechnol. Bioeng. 2017;114: 46-52. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Chemical Sciences, Università degli Studi di Napoli Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, Naples, 4 80126, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Piscitelli
authors.affiliation: Department of Chemical Sciences, Università degli Studi di Napoli Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, Naples, 4 80126, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Pennacchio
authors.affiliation: Department of Chemical Sciences, Università degli Studi di Napoli Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, Naples, 4 80126, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Longobardi
authors.affiliation: Department of Physics, Università degli Studi di Napoli Federico II, Complesso Universitario Monte S. Angelo, Naples, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Velotta
authors.affiliation: Department of Chemical Sciences, Università degli Studi di Napoli Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, Naples, 4 80126, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Giardina
mesh_terms: D000682:Amyloid / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020076:Pleurotus / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012549:Schistosoma japonicum / Q000201:enzymology / Q000235:genetics
chemical_list: D000682:Amyloid; D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase
keywords: amyloid fibrils; biosensors; immobilization; self-assembling proteins