title: Solid-phase handling of hydrophobins: immobilized hydrophobins as a new tool to study lipases.
abstract: Hydrophobins are fungal proteins that self-assemble spontaneously at hydrophilic-hydrophobic interfaces and change the polar nature of the surfaces to which they attach. This attribute can be used to introduce hydrophobic foci on the surface of hydrophilic supports where hydrophobins are attached by covalent binding. In this paper, we report the binding of Pleurotus ostreatus hydrophobins to a hydrophilic matrix (agarose) to construct a support for noncovalent immobilization and activation of lipases from Candida antarctica, Humicola lanuginosa, and Pseudomonas flourescens. Lipase immobilization on agarose-bound hydrophobins proceeded at very low ionic strength and resulted in increased lipase activity and stability. The enzyme could be desorbed from the support using moderate concentrations of Triton X-100, and its enantioselectivity was similar to that of lipases interfacially immobilized on conventional hydrophobic supports. These results suggest that lipase adsorption on hydrophobins follows an "interfacial activation" mechanism; immobilization on hydrophobins offers new possibilities for lipase study and modulation and reveals a new application for fungal hydrophobins.
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis, CSIC, Campus Universidad Autonóma, Cantoblanco, 28049 Madrid, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: Palomo
authors.forename: María M
authors.initials: MM
authors.lastname: Peñas
authors.forename: Gloria
authors.initials: G
authors.lastname: Fernández-Lorente
authors.forename: César
authors.initials: C
authors.lastname: Mateo
authors.forename: Antonio G
authors.initials: AG
authors.lastname: Pisabarro
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernández-Lafuente
authors.forename: Lucía
authors.initials: L
authors.lastname: Ramírez
authors.forename: José M
authors.initials: JM
authors.lastname: Guisán
mesh_terms: D002175:Candida / Q000201:enzymology; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D011551:Pseudomonas fluorescens / Q000201:enzymology
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D008049:Lipase