Estabelecimento de sistema bacteriano de expressão de peptídeos derivados da enzima vegetal RuBisCO

The objective of this research was to establish a bacterial expression system for peptides derived from the in silicosimulated proteolysis of the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase obtained from soybeans, aiming to make a sustainable method for the production of these molecules feasible for future industrial application. Firstly, the Escherichia coli S17-1 strain was made calcium-competent for the propagation of the expression plasmid pET-30a(+), carrying the coding insert for the peptide sequence GSIKAFKEATKVDKVVVLWTALVPR. After plasmid DNA extraction, the material collected was transformed into high-efficient E. coli Rosetta™(DE3)pLysS cells. Rosetta cells carrying the expression plasmid were then induced and peptide production verified through vertical gel electrophoresis, confirming the establishment of a viable expression system for heterologous peptides. Thus, the larger scale production of RuBisCO-derived peptides – along with future purification and activation steps – became possible. In addition, the method set forth herein may also be applied to different peptidic sequences with antimicrobial activity.