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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

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.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.source	SpringerPlus Vol. 2	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	info:eu-repo/semantics/article	es_ES
dc.identifier.doi	10.1186/2193-1801-2-89	es_ES
dc.identifier.idgrec	085670	es_ES