Kinetic model for DBT desulphurization by resting whole cells of Pseudomonas putida CECT5279

Abstract

Bio-desulphurization kinetics of dibenzothiophene (DBT) using Pseudomonas putida CECT 5279, a genetically modified micro-organism (GMO), is studied. A kinetic model describing the 4S route of DBT desulphurization is proposed. Bio-desulphurization experiments have been carried out using resting whole cells of P. putida CECT 5279 obtained at different growth times as biocatalysts. The kinetic equations proposed for each reaction have been previously checked by studying each reaction of the 4S route individually, employing different substrates in different experiments. Finally, simple Michaelis–Menten kinetic equations for the three first reactions catalyzed by two mono-oxygenases (DszC and DszA) and a kinetic equation taking into account competitive inhibition due to product for the final reaction catalyzed by a desulfinase (DszB) have been adopted. DBT has been desulphurized using cells obtained at different growth times (5, 10, 23, 30 and 45 h). The overall kinetic model proposed involving the four reactions of the 4S route was fitted to all the experimental data yielding a set of kinetic parameters able to describe the system evolution. Cell age has influence on the rates of all the reactions: reactions (1), (2) and (3) present maximum rates for cell grown during 30 h, while reaction (4) shows a maximum rate for cells with around 10 h of growth time. However, affinities of each substrate and the inhibition constant of the last reaction are not influenced by the time of growth.