%0 Journal Article
%T A synthetic biology approach to self-regulatory recombinant protein production in Escherichia coli
%A Martin Dragosits
%A Daniel Nicklas
%A Ilias Tagkopoulos
%J Journal of Biological Engineering
%D 2012
%I BioMed Central
%R 10.1186/1754-1611-6-2
%X We present a novel self-regulatory protein production system that couples the control of recombinant protein production with a stress-induced, negative feedback mechanism. The synthetic circuit allows the down-regulation of recombinant protein expression through a stress-induced promoter. We used E. coli as the host organism, since it is widely used in recombinant processes. Our results show that the introduction of the self-regulatory circuit increases the soluble/insoluble ratio of recombinant protein at the expense of total protein yield. To further elucidate the dynamics of the system, we developed a computational model that is in agreement with the observed experimental data, and provides insight on the interplay between protein solubility and yield.Our work introduces the idea of a self-regulatory circuit for recombinant protein products, and paves the way for processes with reduced external control or monitoring needs. It demonstrates that the library of standard biological parts serves as a valuable resource for initial synthetic blocks that needs to be further refined to be successfully applied in practical problems of biotechnological significance. Finally, the development of a predictive model in conjunction with experimental validation facilitates a better understanding of the underlying dynamics and can be used as a guide to experimental design.Recombinant or heterologous protein production (RPP) is an important biotechnological process, with applications that range from catalysis (e.g. washing detergents) and therapeutic use (e.g. antibody production), to protein production for enzymatic characterization and crystallography. Production of human proteins in bacteria dates back to the production of the 14-codon somatostatin gene in Escherichia coli in 1977 [1]. Since then, several hosts have been explored, including other prokaryote species [2], various yeast and fungal species [3], plant, insect, and mammalian cell lines [4]. As there is no universally
%K Synthetic biology
%K Recombinant protein
%K Self-regulatory
%K Escherichia coli
%K Stress promoter
%U http://www.jbioleng.org/content/6/1/2