New Method for Producing Soluble Peptides in E. coli
Author Information
Author(s): Paal Michael, Heel Thomas, Schneider Rainer, Auer Bernhard
Primary Institution: Austrian Center of Biopharmaceutical Technology
Hypothesis
Can a novel Ecotin-Ubiquitin-Tag (ECUT) improve the production of soluble peptides in the periplasm of E. coli?
Conclusion
The study presents a new method for producing authentic soluble peptides in the periplasm of E. coli, which could also be adapted for larger proteins.
Supporting Evidence
- Fusion proteins were expressed in E. coli and translocated into the periplasmic space.
- Under optimal conditions, 18 mg of periplasmic recombinant protein was obtained per gram of dry cell weight.
- Cleavage of target peptides from hybrid proteins was confirmed by mass spectroscopy.
Takeaway
Scientists found a new way to make proteins in bacteria that helps them stay soluble and not clump together, which is important for their function.
Methodology
The study involved expressing fusion proteins in E. coli, followed by purification and cleavage using a deubiquitinating enzyme.
Limitations
The method may be affected by the presence of endogenous ubiquitin-processing enzymes in E. coli, which could reduce product yield.
Digital Object Identifier (DOI)
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