From Biology to Biotechnology: Disulfide Bond Formation in <i>Escherichia coli</i>

Abstract

Disulfide bonds formed between a pair of oxidized cysteines are important to the structural integrity and proper folding of many proteins. Accordingly, Nature has evolved several systems for the genesis and maintenance of such bonds. Beginning with the discovery of protein disulfide isomerase, which provided the first evidence for enzyme-catalyzed disulfide-bond formation, many years of research have resulted in the explication of the complex network of electron transport pathways needed for this process. Herein, we take a historical approach in describing the elucidation of disulfide-bond formation in E. coli. We frame this topic in the context of genome sequencing eras. The first section describes the discovery of eukaryotic protein disulfide isomerase and the subsequent research that followed from the early 1960s to the early 1990s, a time period we have named the pre-genomic sequencing era. The second section details the renaissance in research on disulfide-bond formation in the periplasm of prokaryotes, fueled by bacterial genetic screens and the development of genomic sequencing technology. Accordingly, we have named this section the genomic sequencing era, which ranges from the early 1990s to approximately 2010. The final section outlines the use of bacterial genetic screens to select for new oxidoreductase enzymes and their potential uses in biotechnological and pharmaceutical applications. This era we have dubbed the post-genomic sequencing era, and we envision it to represent the future of research on oxidative folding.

A novel approach for selecting potent peptide inhibitors of the SARS-CoV2-Mpro protease

“…As for peptide (M78), it had 2 cysteines which are highly reactive groups that have the potential to form disulphide bonds not only with the cysteine in the active site of M pro protease but also with each-other in a matter of minutes 36 . Therefore, we suspect that the life of this peptide inhibitor is very short and we did not use the best conditions to detect its activity A weakness in our study is that we only screened a small fraction of all available peptides in our library (1 million clones at each stage of selection).…”

<p>Activity and Effectiveness of Recombinant hEGF Excreted by <em>Escherichia coli</em> BL21 on Wound Healing in Induced Diabetic Mice</p>

“…[21][22][23] The process of protein recombination strongly influences this structure. [23][24][25][26][27][28] One way to produce recombinant hEGF with three disulfide bonds that perfectly fold is by using Escherichia coli (E. coli) with the PelB signal peptide. [29][30][31] The use of E. coli is also beneficial because recombinant hEGF is secreted directly into the periplasmic space and does not mix with cytoplasmic components, making it easy to purify and streamline production costs.…”