Growth of wildtype and mutant E. coli strains in minimal media for optimal production of nucleic acids for preparing labeled nucleotides.
Since RNAs lie at the center of most cellular processes, there is a need for synthesizing large amounts of RNAs made from stable isotope-labeled nucleotides to advance the study of their structure and dynamics by nuclear magnetic resonance (NMR) spectroscopy. A particularly effective means of obtaining labeled nucleotides is to harvest these nucleotides from bacteria grown in defined minimal media supplemented with 15NH4Cl and various carbon sources. Given the high cost of carbon precursors required for labeling nucleic acids for NMR studies, it becomes important to evaluate the optimal growth for commonly used strains under standard minimal media conditions. Such information is lacking. In this study, we characterize the growth for Escherichia coli strains K12, K10zwf, and DL323 in three minimal media with isotopic-labeled carbon sources of acetate, glycerol, and glycerol combined with formate. Of the three media, the LeMaster-Richards and the Studier media outperform the commonly used M9 media and both support optimal growth of E. coli for the production of nucleotides. However, the growth of all three E. coli strains in acetate is reduced almost twofold compared to growth in glycerol. Analysis of the metabolic pathway and previous gene array studies help to explain this differential growth in glycerol and acetate. These studies should benefit efforts to make selective 13C-15N isotopic-labeled nucleotides for synthesizing biologically important RNAs.
Thakur
Chandar S
CS
Department of Chemistry and Biochemistry, Center for Biomolecular Structure & Organization, University of Maryland, 1115 Biomolecular Sciences, Bldg #296, College Park, MD 20742-3360, USA.
Brown
Margaret E
ME
Sama
Jacob N
JN
Jackson
Melantha E
ME
Dayie
T Kwaku
TK
D000085:Acetates / Q000378:metabolism; D002247:Carbon Isotopes / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D005561:Formates / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D007553:Isotope Labeling; D009682:Magnetic Resonance Spectroscopy; D053858:Metabolic Networks and Pathways; D009587:Nitrogen Isotopes / Q000378:metabolism; D009711:Nucleotides / Q000096:biosynthesis*; D012313:RNA / Q000096:biosynthesis*
D000085:Acetates; D002247:Carbon Isotopes; D003470:Culture Media; D005561:Formates; D009587:Nitrogen Isotopes; D009711:Nucleotides; C030544:formic acid; D012313:RNA; D005990:Glycerol