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 mug/ml. Recombinant lipocalins were purified by ion-exchange chromatography from the culture medium. A preliminary NMR characterisation showed that both proteins were correctly folded. (C) 2004 Elsevier B.V. All rights reserved.

Invernizzi, G., Ragona, L., Brocca, S., Pedrazzoli, E., Molinari, H., Morandini, P., et al. (2004). Heterologous expression of bovine and porcine beta-lactoglobulins in Pichia pastoris: towards a comparative functional characterisation. JOURNAL OF BIOTECHNOLOGY, 109(1-2), 169-178 [10.1016/j.jbiotec.2003.10.034].

Heterologous expression of bovine and porcine beta-lactoglobulins in Pichia pastoris: towards a comparative functional characterisation

BROCCA, STEFANIA;LOTTI, MARINA
2004

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 mug/ml. Recombinant lipocalins were purified by ion-exchange chromatography from the culture medium. A preliminary NMR characterisation showed that both proteins were correctly folded. (C) 2004 Elsevier B.V. All rights reserved.
Articolo in rivista - Articolo scientifico
beta-lactoglobulin; expression; Pichia pastoris; NMR
English
8-apr-2004
109
1-2
169
178
none
Invernizzi, G., Ragona, L., Brocca, S., Pedrazzoli, E., Molinari, H., Morandini, P., et al. (2004). Heterologous expression of bovine and porcine beta-lactoglobulins in Pichia pastoris: towards a comparative functional characterisation. JOURNAL OF BIOTECHNOLOGY, 109(1-2), 169-178 [10.1016/j.jbiotec.2003.10.034].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/396
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