Search results for "Recombination-Based Assay"

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

2017

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 expres…

0106 biological sciences0301 basic medicineArabinoseMolecular biologylcsh:MedicineLaccasesBiochemistryBiotecnologia01 natural sciencesSubstrate Specificitylaw.inventionDatabase and Informatics Methodschemistry.chemical_compoundlawRecombinant Protein PurificationCloning MolecularAmineslcsh:Sciencechemistry.chemical_classificationMultidisciplinaryABTSbiologyOrganic CompoundsTemperatureHydrogen-Ion ConcentrationTyramineRecombinant ProteinsEnzymesChemistryRecombination-Based AssayBiochemistryPhysical SciencesRecombinant DNAElectrophoresis Polyacrylamide GelOxidation-ReductionSequence AnalysisResearch ArticleProtein PurificationBioinformaticsTyramineLibrary ScreeningDNA constructionResearch and Analysis Methods03 medical and health sciencesBacterial ProteinsSequence Motif Analysis010608 biotechnologyAmino Acid SequenceBenzothiazolesPediococcus acidilacticiLaccaseMolecular Biology Assays and Analysis TechniquesBase SequenceMolecular massLaccaseOrganic Chemistrylcsh:RChemical CompoundsBiology and Life SciencesProteinsPediococcus acidilacticiSequence Analysis DNAbiology.organism_classificationMolecular biology techniques030104 developmental biologyEnzymechemistryPlasmid ConstructionEnzymologySpectrophotometry Ultravioletlcsh:QSulfonic AcidsEnzimsProteïnesPurification TechniquesPLOS ONE
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