Improved methods of cultivation and production of deuteriated proteins from E. coli strains grown on fully deuteriated minimal medium

摘要: Aims: The aim was to develop reliable and economical protocols for the production of fully deuteriated biomolecules by bacteria. This required preparation deuterium-tolerant bacterial strains an understanding physiological mechanisms acquisition deuterium tolerance. Methods Results: We report here improved methods cultivation Escherichia coli on minimal medium. A multi-stage adaptation protocol developed; this included repeated plating selection colonies resulted in highly cell cultures. Three E. strains, JM109, MRE600 MRE600Rif, were adapted growth succinate is first JM109 being media. showed good, consistent rates capable transformed with plasmids. Expression heterologous proteins these yields consistently high (100–200 mg  l−1). also show that all cells are inherently media. Conclusions: have developed a new three strains. Deuterium-adapted cultures produced good recombinant protein. suggest media, but non-specific mutations enhance tolerance. Thus leads strains. Significance Impact Study: An mechanism media allows strategies development disabled suitable high-level other biomolecules. particular importance nuclear magnetic resonance neutron scattering studies