Construction of a pediocin-producing recombinant lineage from agroindustrial waste

Abstract

A significant effort is underway to develop new antimicrobial compounds for both food and pharmaceutical industry. Bacteriocins are peptides produced by a number of bacteria that harbor bacteriostatic or bactericide effect over pathogenic and deleterious microorganism present in industrial processes. The ones produced by lactic acid bacteria (LABs) present an interest on industry as some of them are probiotic, therefore, acknowledged as safe on this field. In this context, the aim of this project is the maximization of pediocin A production by Pediococcus pentosaceus ET 34 probiotic bacterium through molecular biology tools combined with techniques of scale up improvement on fermentation process. The first step of this project consists in the identification pediocin gene from P. pentosaceus ET 34 and the physical chemical characterization of this bacteriocin. Secondly, the construction of a recombinant strain of P. pentosaceus ET 34 over-expressing heterologously the induction factor and subsequently the comparison of its pediocin productivity with the corresponding wild type via qPCR in order to quantify pediocin transcripts. Finally, we intend to evaluate the effect of different parameters on pediocin productivity in the recombinant strain of P. pentosaceus ET 34. For this purpose, the experiments will be carry out using fractional factorial designs and/or rotational central composite design (RCCD) using agroindustrial waste, such as hydrolyzed sugarcane bagasse with high xylooligosaccharide content. The results obtained will lead to establish the scale up parameters, starting from small scale assays followed by 3L discontinuous bioreactor. (AU)