Engineering bioluminescent Escherichia coli through lux operon expression

Abstract

This study presents a detailed experimental framework for engineering stable bioluminescence in the non-pathogenic Escherichia coli K-12 strain through the in vitro introduction of the lux operon derived from Vibrio fischeri. Using a plasmid-based transformation strategy and heat-shock mediated uptake, the research investigates the feasibility of producing sustained light emission in a genetically tractable bacterial host. Large-scale biomass production was achieved through controlled bioreactor fermentation, followed by chemical competence induction and plasmid transformation. Bioluminescence output was analyzed in relation to cell density measured through impedance-based counting techniques. The work aims to bridge molecular genetics, bioprocess engineering, and quantitative bio-optical analysis, while proposing future applications in biosensing, microbial imaging, and synthetic biology.