Silantes OD-Media

Growing microorganisms on isotopically labeled media is a convenient and effective way to label biomolecules with stable isotopes.
Silantes has developed media for in vivo enrichment of stable isotopes in E. coli and Yeast. Silantes OD-Media are rich growth media optimized to ensure reproducible growth rates, cell density and high yields of heterologous expressed protein.
 
 

 

Silantes Technology

Silantes OD-Media are made from bacterial hydrolysate and contain primarily amino acids, some low MW oligopeptides and almost no carbohydrates. The bacterial strain used is a chemolithoautotrophic organism which grows on inorganic substrates that are isotopically labeled. Due to this technology Silantes rich growth media are very competitively priced.
 
  Free Media-Sample

To test Silantes OD media with your expression system you can obtain a free unlabeled media sample. Usually we send out 200 ml of unlabeled media for testing purposes. For your convenience we invite you to use this Fax-Reply (pdf, 24 kb) or the Silantes contact form for ordering of a media sample.
 

 

 


  fig.2

Added Value

Comparing the expression yield of heterologous protein as well as the time saving factor, Silantes media are often more economical than glucose media.

Consistent results are guaranteed whith Silantes OD-Media. These media yield preset optical densities for different E. coli strains (Fig. 2). In contrast to conventional rich growth media, the growth characteristics and the obtained optical densities of Silantes OD-Media are very similar for labeled and unlabeled media (Fig. 3).

Silantes OD-Media allow researchers to utilize given resources more economically by growing E. coli exactly to the required level. Silantes media provide greater flexibility and predictability. They significantly reduce the costs for expression of heterologous proteins in your laboratory.

 

 

 


  fig.1

Preset OD Values

Silantes OD-Media yield predictable cell densities and exceptional uniform growth curves in shake flask (Fig. 1). As an example, Fig. 2 shows growth curves at OD600 for six commonly used E. coli strains (Fig. 2). (Cell densities are determined at OD600)


OD1 Media (not induced) yield cell densities like a 0,2% glucose-M9 medium
OD2 Media (not induced) yield cell densities like a 0,4% glucose-M9 medium
OD4 to OD5 Media (not induced) yield cell densities as like LB-media

We also supply OD10 Media (concentrate) including salt mix. This concentrate can be diluted to the desired concentration.

 

  fig.3
Convenience

Silantes OD-Media have the same characteristics as conventional LB media: high expression rates and short fermentation times. Silantes OD media are ready-to-use sterile solutions and can be used instantly for the fermentation of bacteria. Silantes media require only the addition of antibiotics and special nutrients needed for the specific expression system.
 
 
  Isotope Enrichment

Silantes OD-Media can be supplied in any combination of 2H, 13C and 15N. The isotopic purity for all isotopes is >98%.
 

Amino Acid Composition

 

 

Amino Acid AA [µmol / l OD1] % of Total
Asp 50,26 20,56
Thr 8,95 3,7
Ser 9,97 4,1
Glu 27,23 11,1
Gly 27,18 11,1
Ala 36,18 14,8
Val 7,29 2,98
Met 4,35 1,78
Ileu 5,15 2,1
Leu 12,85 5,26
Tyr 4,4 1,8
Phe 5,94 2,4
His 23,59 9,65
Lys 8,26 3,38
Arg 5,53 2,3
Pro 7,31 2,99


 

  fig.4
Deuteration

For deuteration with Silantes OD-Media, careful and time consuming adaptation of cells to D2O is not needed for most E. coli strains. Since less generation time is required for the expression of a protein, results can be achieved quicker and more reliable. Combined with the minimal optimization adjustments required and the remarkably high expression yields, deuterated Silantes OD-Media are an interesting and economic alternative to D-glucose minimal D2O media.
Silantes media containing D2O are also ready to use.
  Quality

Quality control tests ensure isotope labeling, sterility and suitability of our media. The preset OD600 value is documented by a quality control certificate provided with each delivery.

  fig.5
Conclusion

Silantes OD-Media are an exceptionally interesting and economic alternative to 13C glucose minimal media because they:

*  are rich growth media
*  are easy to handle
*  have high isotopic purity (>98%)
*  require minimal optimization adjustments
*  ensure high expression yields
*  are very competitively priced

CN-Media "The result was comparable to typical rich growth media. In comparison to glucose-minimal media, the yield on Silantes OD2 was two-times higher; the growth parameters were good. Silantes OD2 media are very convenient to use."
Prof. Sklenar, Masaryk University, Faculty of Science, Lab. of Biomol. Structure and Dynamics, Czech Republic
 
CN-Media "Protein expression can be EASILY optimized in Silantes medium and often the best conditions found in LB standard medium can be used. Silantes saves your time! Silantes media are an excellent opportunity to get labeled samples for MR studies especially if you need a fast preparation for high through-put targets. Today most of the labeled samples of our lab are prepared using Silantes Media."
Dr. Leonardo Gonnelli, CERM, Centro di Risonanze Magnetiche Universita di Firenze, Italy
 
CN-Media "We have used Silantes Expression media for protein expression. The yields were with the first trial superior to our results using minimal medium.I can strongly recommend to use the products by Silantes."
Prof. Harald Schwalbe, Massachusetts Institute of Technology, Department of Chemistry, Cambridge, MA, USA
 
DN-Media "After purification, about 35 mg of pure protein were obtained with Silantes OD2 DN Media. This is even a few milligrams more than we usually get from LB cultures."
Dr. Stefan Prytulla, m-phasys, Tübingen, Germany
 
CN-Media "We compared Silantes unlabeled media with Silantes CN labeled media and found no substantial differences in growth or expression rates. We have enclosed data of a typical growth curve and the resulting protein yield obtained in the same system, using N-labeled M9 medium. Silantes media give extraordinary high results compared to common M9 media."
Frankfurt University, Dr. Klaus Fiebig, Marie-Curie-Str. 11, 6439 Frankfurt/Main, Germany
 

Rich Growth Media for E. coli

Product-N°: Description labeled
(> 98%)
Quantity  
110101402 E. Coli-OD2 D 2H 1000 ml  
110201402 E. Coli-OD2 C 13C 1000 ml  
110301402 E. Coli-OD2 N 15N 1000 ml  
110501402 E. Coli-OD2 DN 2H, 15N 1000 ml  
110601402 E. Coli-OD2 CN 13C, 15N 1000 ml  
110701402 E. Coli-OD2 CDN 2H (> 95%), 13C, 15N 1000 ml  
         
110101401 E. Coli-OD1 D 2H 1000 ml  
110201401 E. Coli-OD1 C 13C 1000 ml  
110301401 E. Coli-OD1 N 15N 1000 ml  
110501401 E. Coli-OD1 DN 2H, 15N 1000 ml  
110601401 E. Coli-OD1 CN 13C, 15N 1000 ml  
110701401 E. Coli-OD1 CDN 2H (> 95%), 13C, 15N 1000 ml  
         
All media are also available as concentrate (OD 10).
         
Please ask for a free sample of our unlabeled media for testing purposes.
         

Silantes Standard Media E. coli

Silantes has developed a new formula of rich growth media that allow a very economical growth of E. coli for in-vivo-labeling with the stable isotope 15N. The Standard-Media are rich growth media that contain the complete set of labeled amino acids, M9 salts and other micronutrients necessary for high expression yields.
A free unlabeled media sample is available on request.

Product-N°: Description labeled
(> 98%)
Quantity  
113301402 Standard-E. Coli-OD2 N 15N 1000 ml  
         
         
Please ask for a free sample of our unlabeled media for testing purposes.

 

Silantes GmbH I Gollierstr. 70 c I 80339 München I Germany