Pesci, LorenzoLorenzoPesciGlueck, Silvia MarinaSilvia MarinaGlueckGurikov, PavelPavelGurikovSmirnova, IrinaIrinaSmirnovaFaber, KurtKurtFaberLiese, AndreasAndreasLiese2019-09-302019-09-302015-02-05FEBS Journal 7 (282): 1334-1345 (2015-04-01)http://hdl.handle.net/11420/3468Microbial decarboxylases, which catalyse the reversible regioselective ortho-carboxylation of phenolic derivatives in anaerobic detoxification pathways, have been studied for their reverse carboxylation activities on electron-rich aromatic substrates. Ortho-hydroxybenzoic acids are important building blocks in the chemical and pharmaceutical industries and are currently produced via the Kolbe-Schmitt process, which requires elevated pressures and temperatures (≥ 5 bar, ≥ 100 °C) and often shows incomplete regioselectivities. In order to resolve bottlenecks in view of preparative-scale applications, we studied the kinetic parameters for 2,6-dihydroxybenzoic acid decarboxylase from Rhizobium sp. in the carboxylation- and decarboxylation-direction using 1,2-dihydroxybenzene (catechol) as starting material. The catalytic properties (Km, Vmax) are correlated with the overall thermodynamic equilibrium via the Haldane equation, according to a reversible random bi-uni mechanism. The model was subsequently verified by comparing experimental results with simulations. This study provides insights into the catalytic behaviour of a nonoxidative aromatic decarboxylase and reveals key limitations (e.g. substrate oxidation, CO2 pressure, enzyme deactivation, low turnover frequency) in view of the employment of this system as a 'green' alternative to the Kolbe-Schmitt processes. Microbial decarboxylases are known to catalyze the reversible regioselective ortho-carboxylation of phenolic derivatives in anaerobic detoxification pathways. In order to get new insights into the catalytic action and to resolve bottlenecks in view applications, we studied the kinetics of 2,6-dihydroxybenzoic acid decarboxylase from Rhizobium sp. in the carboxylation- and decarboxylation-direction, correlating the data according to a reversible random bi-uni mechanism.1742-464X2015713341345Wiley-Blackwellbiocatalytic carboxylationenzyme deactivationkinetic modellingKolbe-Schmitt reactionnonoxidative carboxylationChemieTechnikJournal Article10.1111/febs.13225Other