Biochemical Characterization of Three Heterologous Lactic Acid Bacteria Laccases from Pediococcus, Lactobacillus, and Lactococcus Genus and Their Potential to Degrade Biogenic Amines Using ABTS and Epicatechin as Mediators

Abstract

In this study, we cloned and characterized three bacterial laccases from strains of the species Pediococcus parvulus, Lacticaseibacillus paracasei, and Lactococcus lactis isolated from wine and cheese and evaluated their biogenic amine degradation abilities in the presence/absence of artificial 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) or natural (epicatechin) mediator compounds. Although some recombinant bacterial laccases have been characterized and found to be biological tools for degrading biogenic amines with or without the use of mediators, no prior research has investigated the role of natural mediators, like phenolic substrates found in wine and certain vegetable foods, in the degradation of biogenic amines. The three recombinant bacterial laccases exhibited sigmoidal kinetics and had similar molecular mass but varied in k0.5, kcat, and specific activity toward ABTS. They are acidophilic and have an optimal temperature of 28 °C. However, they exhibit low thermal stability at temperatures higher than 37 °C. The three laccases were capable of degrading dopamine without the use of mediators, while the other amines were not degraded. The presence of ABTS enhanced the degradation of dopamine and tyramine, but the addition of epicatechin did not improve their degradation. This study presents a comparison of the laccases’ biogenic amine-degrading efficiency using different mediators. This is the first time such a comparison has been made.