title: Cloning and characterization of a new laccase from Lactobacillus plantarum J16 CECT 8944 catalyzing biogenic amines degradation.
abstract: In our search for degrading activities of biogenic amines (BAs) in lactic acid bacteria, a protein annotated as laccase enzyme was identified in Lactobacillus plantarum J16 (CECT 8944). In this study, the gene of this new laccase was cloned and heterologously overexpressed in Escherichia coli. The recombinant laccase protein was purified and characterized biochemically. The purified laccase showed characteristic spectroscopic properties of blue multicopper oxidases. The enzyme has a molecular weight of ∼ 62.5 kDa and activity toward typical laccase substrates 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,6-dimethoxyphenol (2,6-DMP). The pH optima on ABTS and 2,6-DMP were 3.5 and 7.0, respectively. Kinetic constants Km and Vmax were of 0.21 mM and 0.54 U/mg for ABTS and 1.67 mM and 0.095 U/mg for 2,6-DMP, respectively. The highest oxidizing activity toward 2,6-DMP was obtained at 60 °C. However, after a preincubation step at 85 °C for 10 min, no residual activity was detected. It has been demonstrated that recombinant L. plantarum laccase oxidizes biogenic amines, mainly tyramine, and thus presents new biotechnological potential for the enzyme in eliminating toxic compounds present in fermented food and beverages.
authors.affiliation: ENOLAB, ERI/ISIC BioTecMed, Universitat de València, Dr. Moliner, 50., E-46100, Burjassot, Valencia, Spain. Sara.Callejon@uv.es.
authors.forename: S
authors.initials: S
authors.lastname: Callejón
authors.affiliation: Departament de Bioquímica i Biologia, Universitat de València, Dr. Moliner, 50., E-46100, Burjassot, Valencia, Spain. Ramon.Sendra@uv.es.
authors.forename: R
authors.initials: R
authors.lastname: Sendra
authors.affiliation: ENOLAB, ERI/ISIC BioTecMed, Universitat de València, Dr. Moliner, 50., E-46100, Burjassot, Valencia, Spain. Sergi.Ferrer@uv.es.
authors.forename: S
authors.initials: S
authors.lastname: Ferrer
authors.affiliation: ENOLAB, ERI/ISIC BioTecMed, Universitat de València, Dr. Moliner, 50., E-46100, Burjassot, Valencia, Spain. Isabel.Pardo@uv.es.
authors.forename: I
authors.initials: I
authors.lastname: Pardo
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D052160:Benzothiazoles / Q000737:chemistry / Q000378:metabolism; D055162:Biocatalysis; D020134:Catalytic Domain; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D042845:Laccase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D048191:Lactiplantibacillus plantarum / Q000737:chemistry* / Q000201:enzymology; D008970:Molecular Weight; D011748:Pyrogallol / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013451:Sulfonic Acids / Q000737:chemistry / Q000378:metabolism; D014439:Tyramine / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D052160:Benzothiazoles; D011994:Recombinant Proteins; D013451:Sulfonic Acids; D011748:Pyrogallol; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; C010120:pyrogallol 1,3-dimethyl ether; D042845:Laccase; D014439:Tyramine
keywords: Biogenic amines; Histamine; Laccase; Lactobacillus plantarum; Putrescine; Tyramine