Concentrate Media

Silantes Concentrate Medium for E.coli and yeasts is a 10-fold concentrated medium obtained from the hydrolysates of stable isotope labeled bacteria fermentations (Cupriavidus nekator). This hydrolysate is a perfect nutrient source for bacteria and yeasts.

The concentrates are available as solution in standard pack sizes or in bulk sizes and can be provided in all 2H, 13C, and 15N combinations.

In our webshop you will find a selection of frequently requested products. Please contact our customer service to see the complete product list or for special requests.

Maximum flexibility

The Silantes Concentrate consists of pure concentrated bacterial hydrolysate. No adjustments have been made to the salt concentrations or other ingredients, as optimization of the ingredients must be made according to the experimental requirements. With the Silantes Concentrate medium, you have maximum flexibility and can dilute and optimize the medium according to your needs.

A manual is included with every delivery, providing you with the recommended amounts of ingredients validated by Silantes for different dilution steps.

Which concentration is best for my experiment?

  • OD600 1 (~ 10-fold dilution): The cell density yields are comparable to 0,2% glucose-M9 media
  • OD6002 (~ 5-fold dilution) is commonly used if inducing the cells at OD600 0.6-0.9 followed by a ~4h incubation. The cell density yields are comparable to 0,4% glucose-M9 media.
    This product is also available as a ready-to-use solution in the webshop.
  • OD600 4 to 5 (~ 2-2.5-fold dilution) is commonly used if growing the cells in a fermenter and/or if growing the cells to higher ODs. The yield cell densities are comparable to LB-media.

Boost the performance of Minimal Media with Silantes Rich Media

Stable isotope-labeled proteins are often expressed in minimal media (or “M9” media). As minimal media contains only essential nutrients required for growth, this can result in slower growth rates and lower biomass compared to rich media, ultimately leading to lower protein yields. This problem can be solved by supplementing the M9 medium with Silantes Rich Media.

Figure 1 demonstrates that 1 % w/v SILEX Powder Medium (or corresponding 1.7 % v/v OD10 Concentrate Medium) added as a supplement to M9 medium increases the initial cell growth rate and the final optical density.

Enhance your stable isotope labeled cell growth and protein yield in minimal media up to 300% in under 10 hours by adding 1% SILEX Powder Medium from Silantes.
Growth of E.coli BL 21 (pRSET-UB) in M9 medium with and without Silantes SILEX medium supplement.

Every delivery includes a manual that provides instructions for use as both “stand-alone” and “supplement to M9”.

Consistent high quality

The 10-fold concentration refers to the performance and implies: When cultivating cells in the undiluted concentrate medium, the cell density reaches a theoretical optical density of OD600 = 10 after 24 hours. The concentrate is standardized – qualitative fluctuations from one batch to the next are prevented.

Medium composition

The medium is based on the bacterial hydrolysate of Cupriavidus necator. Below are the literature values for a number of components.

Component

Composition (g/g DCW) *

Source
Protein 0.680 Srinivasan et al. (2002)
Phospholipid 0.050 Gmeiner et al. (1980)
Cofactors and vitamins 0.030** Ingraham et al., 1983
Cell wall 0.150
Cell wall (Lipopolysaccharide) 0.034 Neidhardt et al. (1996)
Cell wall (Carbohydrate) 0.055 Determined in own study
Cell wall (Peptidoglycan) 0.060 *** Determined in own study

* DCW = Dry Cell Weight. The values are calculated for an average macromolecular composition of Cupriavidus necator H16 in MR minimal medium with D-fructose. The biomass composition was experimentally measured during the exponential growth phase of aerobic batch cultivation (specific growth rate: 0.2 h-1 average of three samples). The molecular weight of one water molecule was subtracted from the molecular weight of each molecule to account for esterification or peptide bonding.

** The assumption is based on the fact that small molecules make up less than 3 % of the dry cell weight.

*** In this study, carbohydrates made up about 5.5 % of the cell wall. The rest was assumed to be peptidoglycan.

The amino acid composition determined for OD2 medium:

Amino Acid AA [ยตmol / l OD2] %
As 131.8 21.5
Thr 17.5 2.9
Ser 24.9 4.1
Glu 75.7 12.4
Gly 68.0 11.1
Ala 77.7 12.7
Val 19.2 3.1
Met 9.1 1.5
Ileu 15.5 2.5
Leu 32.9 5.4
Tyr 10.5 1.7
Phe 17.7 2.9
His 58.2 9.5
Lys 21.0 3.4
Arg 14.4 2.4
Pro 17.5 2.9
611.7 100

References

Relevant manuals:

Use cases of the Silantes OD2 Medium in scientific publications:

  • Mallagaray, A., Creutznacher, R., Dรผlfer, J., Mayer, P. H. O., Grimm, L. L., Orduรฑa, J. M., โ€ฆ Peters, T. (2019). A post-translational modification of human Norovirus capsid protein attenuates glycan binding. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-09251-5
  • Niesteruk, A., Jonker, H. R. A., Richter, C., Linhard, V., Sreeramulu, S., & Schwalbe, H. (2018). The domain architecture of PtkA, the first tyrosine kinase from Mycobacterium tuberculosis, differs from the conventional kinase architecture. Journal of Biological Chemistry, 293(30), 11823โ€“11836. https://doi.org/10.1074/jbc.ra117.000120
  • Pimienta, G. (2004). Structural characterization of a protein/RNA complexโ€ฏ: human TAP/NXF1 protein/retroviral CTE RNA. Heidelberg University Library. https://doi.org/10.11588/HEIDOK.00005077
  • Borland, K., Diesend, J., Ito-Kureha, T., Heissmeyer, V., Hammann, C., Buck, A. H., โ€ฆ Kellner, S. (2019). Production and Application of Stable Isotope-Labeled Internal Standards for RNA Modification Analysis. Genes, 10(1), 26. https://doi.org/10.3390/genes10010026

Use cases of the Silantes SILEX Powder Medium in scientific publications:

  • 2H 13C 15N SILEX Medium for E.coli as substitute to M9: Movellan, K. T., Najbauer, E. E., Pratihar, S., Salvi, M., Giller, K., Becker, S., & Andreas, L. B. (2019). Alpha protons as NMR probes in deuterated proteins. Journal of Biomolecular NMR, 73(1โ€“2), 81โ€“91. https://doi.org/10.1007/s10858-019-00230-y
  • 2H 13C 15N OD2 Medium for E.coli as substitute to M9: Botuyan, M. V. E., Nominรฉ, Y., Yu, X., Juranic, N., Macura, S., Chen, J., & Mer, G. (2004). Structural basis of BACH1 phosphopeptide recognition by BRCA1 tandem BRCT domains. Structure, 12(7), 1137โ€“1146. https://doi.org/10.1016/j.str.2004.06.002
  • 2H 13C 15N OD2 Medium for E.coli as substitute to M9: Lingel, A., Simon, B., Izaurralde, E., & Sattler, M. (2005). The structure of the flock house virus B2 protein, a viral suppressor of RNA interference, shows a novel mode of doubleโ€stranded RNA recognition. EMBO Reports, 6(12), 1149โ€“1155. https://doi.org/10.1038/sj.embor.7400583

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