Optimization of an innovative platform for production of enzymes of marine and freshwater bacteria with biotechnological applications

Energy production from biomass is an attractive alternative to common sources of energy and its study has been growing in recent years. In this context, devices that utilize microorganisms present in wastewaters to produce energy were developed (MFC’s – Microbial Fuel Cells). This technology is still distant from its maximum potential. To optimize it, a deep knowledge of the function of the electricity-producing organisms is needed. Several c-type cytochromes have already been identified as key components in biological energy generation. The production and purification of these proteins is often difficult and therefore, there’s a constant search for optimization of expression systems. The implementation of a new, simpler, expression system (CcHL) in E.coli is an attractive perspective and is, therefore, the aim of this work. For that purpose, the STC gene was inserted on a plasmid previously built for expression of CcHL in E.coli. Cell cultures were grown and two purification steps were conducted. Despite the correct insertion of the gene in the plasmid, NMR spectroscopy and western blotting N-terminal sequence demonstrated the incapacity of E.coli to correctly produce this multi-heme cytochrome. This is probably due to inability of CcHL to assemble more than one heme.

ISCED Categories

The highlighted icons, represent the fields of education (in compliance with ISCED Classification) engaged during this course/programme.

0512 - Biotechnology", "0511 - Biology", "0713 - Offshore and renewable energy