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Escherichia coli K-12 substr. MG1655 Growth Medium: MOPS medium with 0.4% glucose

Summary:
The goals for this medium were to find a convenient and well defined composition that allowed varying all of the elements independently to facilitate isotopic labelling. This required using much less phosphate than in older media, which served roles of pH buffering and chelation of metals like Fe2+. For buffering, MOPS was used instead, with some Tricine added to help chelate Fe2+.

E. coli can use MOPS as a sulfur source if no other sulfur source is added. However, sulfate easily outcompetes MOPS if both are present.

To increase ionic strength and thus optimize the growth rate, NaCl was added. However, it is probably the case that KCl would work just as well, which would allow completely eliminating Na+ from this medium.

Citations: [Neidhardt74]

Recipe Substances:
Substances Concentration Role Comment
β-D-glucopyranose 4 g/l Source of C  
3-(N-morpholino)propanesulfonate 40 mM pH Buffer  
ammonium chloride 9.5 mM Source of N  
ammonium heptamolybdate 3 nM    
borate 400 nM    
calcium chloride 0.5 µM    
cobalt chloride 30.0 nM    
copper sulfate 10 nM    
dipotassium phosphate 1.32 mM Source of P  
iron sulfate 10 µM    
manganese chloride 80 nM   Mn source
MgCl2 528 µM    
potassium sulfate 276 µM Source of S  
sodium chloride 50 mM    
tricine 4 mM pH Buffer  
zinc sulfate 10 nM    
Composition:
Constituents Concentration
chloride 60.557 mM
Na+ 50 mM
3-(N-morpholino)propanesulfonate 40 mM
borate 24.732 mM
β-D-glucopyranose 22.203 mM
ammonium 9.5 mM
tricine 4 mM
K+ 3.192 mM
phosphate 1.32 mM
Mg2+ 528 µM
sulfate 286.02 µM
Fe2+ 10 µM
Ca2+ 500 nM
Mn2+ 80 nM
Co2+ 30.0 nM
Cu2+ 10 nM
Zn2+ 10 nM
heptamolybdate 3 nM

pH: 7.2

Osmolarity (approximate, computed from constituents): 0.22 Osm/L

Wildtype growth observations:

T (°C) O2 Growth?
37 Aerobic Yes

15 single gene knockouts exhibit no growth (aerobic):
argH , aroA , carA , carB , guaA , ppc , proB , proC , purA , purC , purE , purH , serC , thrC , thyA (T=37°C) [Feist07, Baba06, Comment 1]

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3736 single gene knockouts exhibit growth (aerobic):


aaeA , aaeB , aaeR , aaeX , aat , abgA , abgB , abgR , abgT , abpA , abpB , abrB , aceK , acrA , acrB , acrD , acrR , acrZ , ada , adeP , adeQ , adiY , aegA , aer , aes , afuB , afuC , agaA , agaB , agaC , agaD , agaI , agaR , agaS , agaV , agaW , ahpC , ahpF , ahr , aidB (T=37°C) [Baba06, Comment 2] + 3696 more...
[Show All]

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Genes with inconsistent growth observations:

Knockout Growth? T (°C) O2 Growth Observations
aceFIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
argAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
argBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
argCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
argEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
argGIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
aroBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
aroCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
aroEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
atpBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
atpCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
atpEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
bioAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
bioBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
bioDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
bioFIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysGIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysHIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysIIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysJIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
cysNIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
glnAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
gltAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
glyAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
guaBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisFIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisGIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisHIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
hisIIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
icdIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ilvAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ilvCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ilvDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ilvEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
iscSIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
leuAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
leuBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
leuCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
leuDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
lpdIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
lysAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
metAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
metBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
metCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
metEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
metFIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
metLIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
nadAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
nadBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
nadCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pabAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pabBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
panBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
panCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
panDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pdxAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pdxBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pdxHIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pdxJIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pfkAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pheAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
proAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ptsHIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ptsIIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
purDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
purFIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
purKIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
purLIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
purMIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pyrBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pyrCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pyrDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pyrEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
pyrFIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
serAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
serBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
thrAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
thrBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
tnaBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
trpAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
trpBIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
trpCIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
trpDIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
trpEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
tyrAIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]
ubiEIndeterminate 37 Aerobic
Yes [Baba06, Comment 2]
No [Feist07, Comment 4]

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References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Neidhardt74: Neidhardt FC, Bloch PL, Smith DF (1974). "Culture medium for enterobacteria." J Bacteriol 119(3);736-47. PMID: 4604283


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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