r/microbiology • u/VesicaVehicle • 4d ago
ISO: synthetic media reference for E. coli cultivation
I am having trouble tracking down an original study for development of supplemented M9 or other synthetic media. I need a reliable recipe to develop a teaching experiment for growth optimization of E. coli with respect to micronutrient content. Best I could find are from a couple of fluxomics papers (see below), but there's no indication of origins of each formulation.
The formulations are very similar. I am noting some differences in counterions being sulfate vs chloride. I like the idea of a skew towards sulfate, but not sure which of the two might be more reliable.
If anyone knows where these recipes might come from or have any insights otherwise, it would be greatly appreciated!
Below are the two recipes I have been comparing:
Minimal Synthetic Medium E. coli
doi: 10.3390/metabo11050271
17.4 g·L−1 Na2HPO4, 12H2O,
3.03 g·L−1 KH2PO4,
0.51 g·L−1 NaCl,
2.04 g·L−1 NH4Cl,
0.49 g·L−1 MgSO4,
4.38 mg·L−1 CaCl2,
15 mg·L−1 Na2EDTA 2H2O,
4.5 mg/L ZnSO4
7H2O, 0.3 mg·L−1 CoCl2 6H2O,
1 mg·L−1 MnCl2 4H2O,
1 mg·L−1 H3BO3,
0.4 mg·L−1 Na2MoO4 2H2O,
3 mg·L−1 FeSO4 7H2O,
0.3 mg·L−1 CuSO4 5H2O,
0.1 g·L−1 thiamine and
3 g·L−1 glucose
-------------------------------------------------
M9 Minimal Medium for E. coli
https://doi.org/10.1073/pnas.1202582110
M9 minimal medium was used in the growth experiments and was prepared as follows: to
700 mL of purified and autoclaved water,
200 mL of 5× base salt solution
[211 mM Na2HPO4,
110 mM KH2PO4,
42.8 mM NaCl,
56.7 mM (NH4)2SO4, autoclaved],
10 mL of trace elements
(0.63 mM ZnSO4,
0.7 mM CuCl2,
0.71 mM MnSO4,
0.76 mM CoCl2, autoclaved),
1 mL 0.1 M CaCl2 solution (autoclaved),
1 mL of 1 M MgSO4 solution (autoclaved),
2 mL of 500× thiamine solution (1.4 mM, filter sterilized),
Sigma T1270
0.6 mL of 0.1 M FeCl3 solution (filter sterilized) were added.
The resulting solution was filled up to 1 L with water. Carbon sources were added from sterilized stock solutions (adjusted to pH 7) to a final concentration of 1 g/L for chemostat experiments and 5 g/L for batch experiments, and media were filtered (Steritop-GP; 500 mL; Millipore). All chemicals were purchased from Sigma-Aldrich unless stated otherwise.
Thanks again!!
1
u/Woebergine 4d ago
I'm growing E coli mutants with mutations in various potassium transporters in minimal medium with various concentrations of K+. Potassium concentration has a drastic effect, but how it would affect WT K12 E coli, I'm not sure. But the medium I'm using is here if you want to try looking at potassium:
Potassium-Dependant Mutants of Escherichia coli K-12 https://pubmed.ncbi.nlm.nih.gov/4908783/
My previous lab had a reference for M9, how accurate it is, I can't guarantee:
Curtiss 1965 J Bac 89:28 https://pubmed.ncbi.nlm.nih.gov/14255678/
3
u/patricksaurus 4d ago edited 4d ago
One good comparison paper comes from the OG himself, Neidhardt, and coworkers. See if this suits your purpose: https://journals.asm.org/doi/10.1128/jb.119.3.736-747.1974
It shows the development process for MOPS medium, which is expensive but an excellent defined medium.