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. 2020 Jan 13;10:191. doi: 10.1038/s41598-019-57051-0

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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LCMS based analysis of metabolic pool sizes in PCC 11802 and 11801 under elevated CO₂ conditions. The bar graphs represent the ratio of metabolite pools at 1% CO₂/ambient (0.04%) CO₂ in PCC 11802 and 11801. The metabolites represented in the figure show the difference between PCC 11802 and 11801 at a p-value < 0.01 using the t-test with the exception of phosphoenolpyruvate (PEP), acetyl coenzyme A (Ac-CoA) and ADP-glucose (ADPG) which show p-values of 0.02, 0.09 and 0.5 respectively. The relative metabolite pool sizes in each condition were estimated by extracting the metabolites as described previously and mixing with an extract of PCC 11801 which is fully labeled with isotopic ¹³C carbon. High-resolution LCMS data was collected by achieving separation of metabolites using a reverse phase, ion-pairing chromatography method. The ratio of areas of the ¹²C monoisotopic peak to the respective ¹³C monoisotopic peak was quantified for each metabolite, which served as a measure of relative pool size in a given condition. Abbreviations used for metabolites, ADPG: ADP-glucose, Ac-CoA: acetyl coenzyme A, AKG: alpha-ketoglutaric acid, ASP: aspartic acid, CIT: citric acid, DHAP: dihydroxyacetone phosphate, FBP: fructose 1,6 bisphosphate, F6P: fructose-6-phosphate, GAP: glyceraldehyde-3-phosphate, GLU: glutamate, G1P: glucose-1-phosphate, G6P: glucose-6-phosphate, ISO-CIT: isocitric acid, MAL: malic acid, OAA: oxaloacetic acid, PEP: phosphoenolpyruvate, PYR: pyruvic acid, RuBP: ribulose 1,5 bisphosphate, Ru5P: ribulose-5-phosphate, R5P: ribulose-5-phosphate, SBP: sedoheptulose 1,7 bisphosphate, SSA: succinyl semialdehyde, SUC: sucrose, SUCC: succinate, SUC-6-P: sucrose-6-phosphate, S7P: sedoheptulose-7-bisphosphate, UDPG: UDP-glucose, 2PGA: 2-phosphoglyceric acid, and 3PGA: 3-phosphoglyceric acid.