EnPresso® B Growth Systems (E. coli protein data)

EnPresso(TM) B Growth Systems have been used to express a variety of protein types in a variety of different cells. The data on the chart below summarizes the proteins and cells used by our customers. Scroll below the chart to see real protein expression data submitted by EnPresso(TM) B users.

 

Examples of recombinant proteins expressed in EnPresso B growth systems Bacterial strains grown successfully
Protein type Target protein Host strain
Protein domain CnaBC2, D5 and E3 domains – CnaB domains from Sdr family proteins (surface adhesins); SdrE proteins E. coli BL21 (DE3) – T1R
Proteases Extracellular metalloprotease of Clostridium difficile: 1.  Unknown hypothetical Zinc metallopeptidase CD630_28300: named Zmb1 and 2. FliD and 3. CD630_23650 E. coli T7 Express
Human proteases Novel chimeric-truncated form of tissue-type plasminogen activator (t-PA – a serine protease) with improved fibrin affinity E. coli BL21 (DE3)  with F– dcm ompT hsdS (rB– mB–) gal genotype
Human lipoxygenases Synonymous genetic variations in four different human lipoxygenase genes (ALOX5, ALOX12, ALOX15, and ALOX15B) E. coli BL21(DE3) pLysS
RNA-binding proteins Nucleolin
EF-hand domain-containing protein D2 (EFHD2)
Serine/arginine-rich splicing factor 1 (SRSF1)
Splicing factor U2AF (U2AF653)
E. coli BL21 (DE3) – T1R
Non-ribosomal peptide synthetases 654 kDa heterodimeric valinomycin synthetase (VlmSyn formed from Vlm1 and Vlm2) from Streptomyces tsusimaensis E. coli BL21 Gold
Single-domain antibodies (sdAbs) Single-domain antibodies (sdAbs) E. coli Shuffle® T7 Express
E. coli BL21(DE3)
Fab antibody fragments Fab antibody fragments E. coli RV308
E. coli BL21
Esterase Novel esterase (Est22) derived from an acidic leachate environment not specified
Cyclophilins Human cyclophilin-A E. coli C41[pEPPF:hCypA]
Dehalogenases Haloalkane dehalogenase: Spur
Haloalkane dehalogenase: LinB
Haloalkane dehalogenase: DbjA
Haloalkane dehalogenase: DhaA
E. coli BL21(DE3)
Dehydrogenases Lactobacillus alcohol dehydrogenase E. coli RB791 [F-, IN(rrnD-rrnE1), l-, lacIqL8]
Biotin-binding proteins Avidin Tes29 not specified
Biotin-binding proteins Bradavidin with C-terminal cysteine not specified
not specified not specified E. coli Rosetta DE3
not specified not specified E. coli BL21-A1

 

Results of Bacterial Culture (Proteins for Research)

Low yield, poor solubility, low activity – just some of the challenges that waste time and effort when producing recombinant proteins for research purposes. EnPresso B growth systems overcome these issues, providing cultivation conditions for bacteria, such as E. coli, that increase yield and provide ’fit for purpose’ proteins. Examples shown here are from independent laboratories – because results (and costs) count.



>30-fold increase in active enzyme with > 3.3-fold increase in specific activity
Results courtesy of Lukáš Chrást, Masaryk University, Czech Republic

Researchers compared EnPresso B with a commercially-available LB growth media.  They recommend EnPresso B for routine E. coli cultures over-expressing recombinant proteins.  They achieved these highlights:

  • Higher cell density (15 to 30-fold) and higher specific activity (1.5 to 3.3-fold)
  • Significant improvement in yield of active enzyme per culture volume
  • The same expression profile during a 24h induction time





66-fold increase in yield of a toxic protein

Result courtesy of Thomas Horn, Charité, Berlin, Germany

Researchers compared EnPresso B with a commercially-available LB growth media.  Their work with EnPresso B gave the following results:

  • Significantly reduced labor in downstream processing with smaller volumes and fewer shake flasks
  • Greatly improved yield and reproducibility
  • Saved costs by reducing IPTG consumption





20-fold increase in final, purified protein yield

Result courtesy of Dr. Martin Wear, University of Edinburgh

Researchers compared EnPresso B with a commercially-available LB growth media. The data from 50mL cultures in shake flasks shows yield of the protein human cyclophilin-A in E coli cells.





>15-fold increase in yield of fusion peptides

Results courtesy of Michael Crampton, CSIR Pretoria, South Africa

Researchers compared EnPresso B with a commercially-available LB growth media.  The data from shake flasks shows increased yield of two different enzymes produced.





5-fold increase in volumetric activity

Result courtesy of major pharmaceutical company

Researchers compared EnPresso B with a commercially-available LB  and HTMC auto-induction growth media.  Their work with EnPresso B gave the following results:

  • Protein expression level per g biomass comparable to LB
  • Reduced consumption of media and antibiotics
  • Saved shaker space to increase throughput
  • Eliminated time-consuming O.D. measurements





100-fold increase in yield of active, soluble protein
Ukkonen, K., et al. 2011. Microb. Cell. Fact. 10:107.

Researchers compared EnPresso B with a commercially-available LB growth media. The data is shown for lactobacillus alcohol dehydrogenase produced in E. coli.

 

Materials

     

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