EnPresso® B Defined Nitrogen-free (E. coli protein data)

Results of Bacterial Culture (Proteins for NMR Analysis)

Poor growth, low levels of expression and insufficient labeled protein are the most frequent challenges faced when using minimal medium to prepare proteins for structural characterization by NMR analysis. EnPresso B Defined Nitrogen-free is a pre-sterilized, chemically-defined growth system that increases the yield of 15N-labeled proteins. Examples shown here are from independent laboratories – because results count.



64-fold increase in yield of
15N-labeled protein
Result courtesy of Julia Smith, Senior Scientist and Dr. Ben Davis, Vernalis, UK

Researchers compared EnPresso B Defined Nitrogen-free with M9 growth medium and presented results at ELRIG Advances in Recombinant Protein Technology.  Highlights of the results include:

  • protein yield of 160 mg/L from EnPresso compared to 2.5 mg/L from M9.
  • high level of 15N incorporation
  • 2D 15N-1H HSQC NMR spectrum showed no significant difference (compared to M9 protein production)





100mg/L of a toxic cytokine fragment – no protein when using M9

Result courtesy of Professor Birthe Kragelund, Dept. of Biology, University of Copenhagen, Denmark

Researchers compared EnPresso B Defined Nitrogen-free  with LB and M9 growth media.  Researchers were able to produce ¹⁵N-labeled cytokine fragments – something that was previously impossible using M9.

      



More labeled protein produced per cell weight (biomass)

Result courtesy of Dr. Anne Diehl, Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany

Researchers compared EnPresso B Defined Nitrogen-free  with a modified M9 growth media.  Highlights from their results:

  • increase in product-specific yield made purification much easier
  • surprisingly high yields
  • degree of labeling and the spectra were identical to previous experiments





5-fold increase in yield of purified 15
N-labeled SPSB2
Result courtesy of Dr. Eleanor Leung, Faculty of Pharmacy and Pharmaceutical Sciences , Monash Institute of Pharmaceutical Sciences, Australia

Researchers compared EnPresso B Defined Nitrogen-free with M9 growth media. Researchers were pleased with the results and noted that the 2D 15N-1H HSQC NMR spectrum was identical to the previously reported spectrum.

   



5-fold increase in yield of three purified protein variants

Result courtesy of major pharmaceutical company, Italy

Researchers compared EnPresso B Defined Nitrogen-free with M9 growth media. Researchers note that the quality of the proteins was confirmed by their ability to bind to their co-factors.

Materials

     
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