SILEX and ECO Powder Media

Silantes SILEX and ECO media are powder media obtained from the Silantes OD2 medium.

The SILEX Powder Medium is a lyophilisate of the Silantes OD2 medium. It combines the performance of the OD2 medium with a good shelf life.

SILEX and ECO Powder Media for E. coli and yeastThe ECO Powder Medium is also a lyophilisate of the OD2 medium, but with a higher variation in the occurrence of low molecular weight oligopeptides. This makes the performance of the ECO medium less reliable compared to the SILEX medium. The ECO powder medium is particularly suitable for validating new systems for isotope-labeled growth at low cost.

The Silantes Powder Media are available in standard pack sizes or in bulk sizes and can be provided in all 2H, 13C and 15N combinations.

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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”.

No Addition of 13C-Glucose Needed

As the raw material 13C-Glucose becomes more and more scarce and thus expensive, it is important to explore alternative options to these chemicals, especially for academic applications. One such alternative is the use of stable isotope-labeled complex media from Silantes: All Silantes cell growth media are prepared from uniformly labeled Cupriavidus necator bacterial cell hydrolysate. This isotopically labeled biomass is a perfect nutrient medium for bacteria and yeasts, eliminating the need for expensive 13C glucose if used as a stand-alone medium.

Find out more about the 13C-glucose price dynamics in the blog article โ€œWhy has the stable isotope price skyrocketed?โ€

Maximum cost efficiency

The high performance of Silantes complex media combined with the economic benefits of powder media offer you maximum cost efficiency. This excellent property also extends to cell growth under deuterated conditions:

Figure 2 shows the growth curve of E.coli in 2H-labeled Silantes OD2 Solution Medium for E.coli and in 1 % (=10g/L) Silantes SILEX Powder Medium for E.coli in D2O. Both media have the same excellent performance in D2O and exceed OD600 2.5 in less than 24 hours.

Figure 2: Growth of E.coli in 2H-OD2 media solution and in SILEX powder media in D2O

In order to evaluate the cost-effectiveness of Silantes Rich media for your project, the respective costs of a corresponding minimal medium in relation to the protein yield can be used for comparison. For the Silantes SILEX and ECO media the following applies:

  • Price Silantes SILEX Medium = Price minimal medium + 15%
    > If you achieve a 15% increase in your protein yield by using the Silantes SILEX medium, you save money by using the SILEX medium.
  • Price Silantes ECO Medium = Price minimum medium
    > If you achieve the same protein yield by using the Silantes ECO medium, you save money by using the ECO medium.

We have validated that expression can be increased by over 200% for a number of proteins using Silantes Rich Medium compared to Minimal Medium.

To validate Silantes Rich Media for your system, contact us for a free sample

High Standards in Quality Control

Silantes media are tested for reproducibility of the fermentation results. Moreover, each batch is adjusted to yield the same cell density. The isotopic enrichment of > 98 % is validated by mass spectrometry and the biological competence by growth tests. All test results are documented in a quality certificate, which is provided with every delivery.

Medium composition

The medium is based on a Cupriavidus necator bacterial hydrolysate. 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 (equivalent to 10g SILEX powder):

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

Related products

Silantes also offers stable isotope labeled reagents for M9 media such as 13C-glucose, D2O and 15NH4Cl.

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

Relevant blog articles:


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