Stanzione, Ilaria (2022) FUNGAL PROTEIC BIOSURFACTANTS FOR THE DEVELOPMENT OF BIOSENSING PLATFORMS. [Tesi di dottorato]

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Collection description

Protein biosurfatants produced by marine and terrestrial filamentous fungi represent the best example of biomolecules that can be widely used as a replacement of their synthetic counterparts, thanks to their low toxicity and biodegradability. Among them, hydrophobins and ceratoplatanins are proteins able to self-assemble at hydrophobic/hydrophilic interfaces forming amphiphilic layers. It is well known that the hydrophobins are involved in the fungal growth thanks to their capability to reduce the surface tension of the air/water interface, instead, while the ceratoplatanins can act both as virulence factor and as elicitors. Moreover, the amphiphilic nature of both classes of proteins makes them of great interest in many fields. For this reason, one of the purposes of the present PhD project is the isolation and characterization of protein biosurfactants from marine fungal strains. The isolated protein from Penicillium chrysogenum has been characterized both as biosurfactant and bioemulsifier, indeed its capabilities to stabilize emulsion and to reduce the surface tension have been verified. Moreover, it has been proved that two marine fungi, Paradendryphiela salina and Talaromyces pinophilus grown on seaweed wastes, are able to produce both cerato-platantins and hydrophobins. Another hydrophobin deeply characterized and exploited in my research group, is Vmh2, produced by the edible fungus P. ostreatus. This protein can form protein layers on several surfaces changing their physical-chemical properties and allowing the attachment of other biomolecules in their active form. Thus, the exploitation of Vmh2 in the diagnosis of cultural heritage and in biosensing field has been under investigation in this PhD project. The functionalization of cellulose acetate sheets with Vmh2 has been employed to immobilize proteolytic enzymes, trypsin and PNGaseF, developing a portable, easy-to-use and non-invasive kit for the identification of ancient proteins on cultural heritage objects. Furthermore, in biosensing field the same hydrophobin has been genetically fused to two Single chain Fragment variables of two antibodies able to bind algal marine neurotoxins, and to a laccase enzyme that can oxidize aromatic and phenolic compounds. The recognition ability of chimeric proteins has been coupled with the interesting characteristics of nanomaterials, such as graphene, carbon nanotubes and magnetic beads. The functionalized 2D-materials with both chimeric proteins have been exploited to develop electrochemical and optical biosensors for the detection of the analytes mentioned above.

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