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Production of soluble eukaryotic recombinant proteins in E. coli is favoured in early log-phase cultures induced at low temperature

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Production of soluble eukaryotic recombinant proteins in E. coli is favoured in early log-phase cultures induced at low temperature

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dc.contributor.author San Miguel Díez, Teresa es_ES
dc.contributor.author Pérez-Bermúdez, Pedro es_ES
dc.contributor.author Gavidia, Isabel es_ES
dc.date.accessioned 2015-06-18T12:27:01Z
dc.date.available 2015-06-18T12:27:01Z
dc.date.issued 2013 es_ES
dc.identifier.citation SpringerPlus Vol. 2 es_ES
dc.identifier.uri http://hdl.handle.net/10550/44460
dc.description.abstract BackgroundProducing recombinant plant proteins expressed in Escherichia coli produce in high yields and in a soluble and functional form can be difficult. Under overexpression conditions, proteins frequently accumulate as insoluble aggregates (inclusion bodies) within the producing bacteria. We evaluated how the initial culture density, temperature and duration of the expression stage affect the production of some eukaryotic enzymes in E. coli.FindingsA high yield of active soluble proteins was obtained by combining early-log phase cultures and low temperatures for protein induction. When IPTG was added at OD600 = 0.1 and cultures were maintained at 4°C for 48-72 h, the soluble protein yield was 3 fold higher than that obtained in the mid-log phase (OD600 = 0.6). Besides, the target protein expression increased and the endogenous bacterial proteins reduced, thus making the protein purification process easier and more efficient.ConclusionsThe protocol can be widely applied to proteins with a heterologous expression which was limited by loss of activity at high temperatures or by low soluble recombinant protein yield. es_ES
dc.subject Early log phase es_ES
dc.subject Functional proteins es_ES
dc.subject Low temperature es_ES
dc.subject Soluble recombinant proteins es_ES
dc.title Production of soluble eukaryotic recombinant proteins in E. coli is favoured in early log-phase cultures induced at low temperature es_ES
dc.type journal article es_ES
dc.identifier.doi 10.1186/2193-1801-2-89 es_ES
dc.identifier.idgrec 085670 es_ES

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