Journal: bioRxiv

Article Title: Autotrophic growth of E. coli is achieved by a small number of genetic changes

doi: 10.1101/2023.06.03.543535

Figure Lengend Snippet: (A ) left: growth curve of an isolated evolved clone vs the engineered compact strain in liquid M9 minimal media supplemented with 45mM sodium formate and sparged with a gas mixture of 10% CO 2 , 5% oxygen, and 85% nitrogen. right: calculated growth rate using linear regression of the engineered compact strain (light blue) and evolved strain (orange), the calculated doubling time at the given conditions is about 24h (μ ≈ 0.03h -1 ). Growth was carried out in triplicates (n=3) in a Spark plate reader with gas control, dark grey area corresponds to standard deviation (150ul culture + 50ul mineral oil, to prevent evaporation). (B) The fractional contribution of 13 CO 2 to various protein-bound amino acids of the compact autotrophic strain after ≈4 doublings on 13 CO 2 and 13 C labeled formate (light blue, mean of n=3) reached the expected 13 C labeling fraction of the biosynthesized amino acids. When grown in naturally labeled CO 2 and 13 C labeled formate (dark blue, n=1, technical triplicate) the 13 C content dropped to close to natural abundance. Experiments with 13 CO 2 as the substrate were carried out in air-tight (i.e., sealed) growth vessels.

Article Snippet: For the measurements of intracellular glucose-6-P and fructose-6-P, metabolic extracts were 10x concentrated using a vacuum evaporation (Eppendorf Concentrator plus) and resuspended in 40:40:20 (v-%) acetonitrile/methanol/water extraction solution.

Techniques: Isolation, Control, Standard Deviation, Evaporation, Labeling