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Recombinant laccase from Pediococcus acidilactici CECT 5930 with ability to degrade tyramine

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Recombinant laccase from Pediococcus acidilactici CECT 5930 with ability to degrade tyramine

dc.contributor.author	Callejón Salinas, Sara
dc.contributor.author	Sendra Pérez, Ramón
dc.contributor.author	Ferrer Soler, Sergi
dc.contributor.author	Pardo Cubillos, Isabel
dc.date.accessioned	2017-11-20T15:03:47Z
dc.date.available	2017-11-20T15:03:47Z
dc.date.issued	2017
dc.identifier.citation	Callejón Salinas, Sara Sendra Pérez, Ramón Ferrer Soler, Sergi Pardo Cubillos, Isabel 2017 Recombinant laccase from Pediococcus acidilactici CECT 5930 with ability to degrade tyramine Plos One 12 10 e0186019
dc.identifier.uri	http://hdl.handle.net/10550/63168
dc.description.abstract	Biogenic amines degradation by bacterial laccases is little known, so we have cloned and heterologously expressed, in E. coli, a new laccase from Pediococcus acidilactici CECT 5930 (Lpa5930), a lactic acid bacterium commonly found in foods able to degrade tyramine. The recombinant enzyme has been characterized by physical and biochemical assays. Here we report the optimization of expression and purification procedures of this laccase. DNA encoding sequence of laccase from P. acidilactici was amplified by PCR and cloned into the expression plasmid pET28a for induction by isopropyl-β-D-thiogalactoipyranoside. Protein expression was performed in E. coli BL21(DE3) harboring pGro7 plasmid expressing a chaperone folding assistant induced by arabinose. Purification was performed by column metal-chelating chromatography on Ni-NTA-agarose. The laccase enzyme obtained has an apparent molecular mass of ∼60 kDa, an optimum temperature activity toward 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) of 28°C, and was quickly inactivated at temperatures higher than 70°C. The apparent Km value for ABTS was 1.7 mM and the Vmax obtained was 24 U/mg. In addition to ABTS, recombinant Lpa5930 laccase degraded the biogenic amine tyramine at pH 9.5 and pH 4.0 with or without ABTS as a mediator. Tyramine degradation by laccases could solve the problems generated in food due to the presence of this toxic compound.
dc.language.iso	eng
dc.relation.ispartof	Plos One, 2017, vol. 12, num. 10, p. e0186019
dc.subject	Enzims
dc.subject	Proteïnes
dc.subject	Biotecnologia
dc.title	Recombinant laccase from Pediococcus acidilactici CECT 5930 with ability to degrade tyramine
dc.type	journal article	es_ES
dc.date.updated	2017-11-20T15:03:47Z
dc.identifier.doi	10.1371/journal.pone.0186019
dc.identifier.idgrec	121089
dc.rights.accessRights	open access	es_ES