title: Heterologous expression of bovine and porcine beta-lactoglobulins in Pichia pastoris: towards a comparative functional characterisation.
abstract: Bovine and porcine beta-lactoglobulins were cloned and expressed in host cells with the aim of developing the tools necessary for their structural, functional and conformational characterisation by NMR techniques. Both lipocalins were expressed in Pichia pastoris, where the use of a constitutive promoter turned out to allow the highest productivity. The yield of recombinant proteins was further improved through multiple integration of the encoding genes and by increasing aeration of the transformed cultures. Both proteins were obtained in the culture medium at the concentration of 200 microg/ml. Recombinant lipocalins were purified by ion-exchange chromatography from the culture medium. A preliminary NMR characterisation showed that both proteins were correctly folded.
authors.affiliation: Dipartimento di Biotecnologie e Bioscienze, Università degli Studi di Milano-Bicocca, Piazza della Scienza 2, Milan, Italy.
authors.forename: Gaetano
authors.initials: G
authors.lastname: Invernizzi
authors.forename: Laura
authors.initials: L
authors.lastname: Ragona
authors.forename: Stefania
authors.initials: S
authors.lastname: Brocca
authors.forename: Eros
authors.initials: E
authors.lastname: Pedrazzoli
authors.forename: Henriette
authors.initials: H
authors.lastname: Molinari
authors.forename: Piero
authors.initials: P
authors.lastname: Morandini
authors.forename: Maddalena
authors.initials: M
authors.lastname: Catalano
authors.forename: Marina
authors.initials: M
authors.lastname: Lotti
mesh_terms: D000818:Animals; D002417:Cattle / Q000235:genetics; D003001:Cloning, Molecular; D015870:Gene Expression; D007782:Lactoglobulins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D010843:Pichia / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D013552:Swine / Q000235:genetics
chemical_list: D007782:Lactoglobulins; D011994:Recombinant Proteins