General principle of the photo-electro-autotrophic <i>E</i>. <i>coli</i>.

<p>Key components are a carbon fixation cycle, a mechanism to take up external electrons and the proton-pumping rhodopsins photosystem.</p

Synthetic carbon fixation pathways: PyrS-PyrC-Glx bicycle (red and black) and the PyrS-PEPC-Glx bicycle (blue and black).

<p>Enzymes involved in the bicycles: 1) pyruvate synthase (oxygen sensitive), 2) pyruvate water dikinase, 3) phosphoenolpyruvate carboxylase, 4) pyruvate carboxylase, 5) malate dehydrogenase, 6) malate thiokinase, 7) malyl-CoA lyase, 8) isocitrate lyase, 9) aconitate hydratase, 10) ATP citrate lyase, 11) fumarate hydratase and 12) fumarate reductase.</p

Additional file 1 of Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO

Additional file 1: Table S1. List of primers used in this study. Table S2. Sequences of genes expressed in this study. Figure S1. Image of Sanger sequencing results showed the nature of the in-frame deletion in the pta gene (CAETHG_3358). Figure S2. Image of Sanger sequencing results showed the nature of the in-frame deletion in the Ald subunit of the adhE1 gene (CAETHG_3747). Figure S3. Change in CO headspace pressure for C. autoethanogenum strains carrying plasmids for AAT expression on CO as main carbon source. Figure S4. Screening of ethyl acetate production by C. autoethanogenum strains carrying plasmids for AAT expression on 40 mM fructose. Figure S4. Investigating growth of C. autoethanogenum on CO and ethyl acetate. Figure S5. Investigating ethyl acetate degradation by C. autoethanogenum grown on CO. Figure S6. Eat1 in vivo alcoholysis assay for C. autoethanogenum and E. coli. Figure S7. Investigating the effects of ethanol supplementation on growth of C. autoethanogenum [PThl-Atf1] grown on CO. Figure S8. Investigating the effects of ethanol supplementation on ethyl acetate production by C. autoethanogenum [PThl-Atf1] grown on C

General characteristics of the analyzed carbon fixation pathways.

<p>General characteristics of the analyzed carbon fixation pathways.</p

Integration of different carbon fixation pathways in the <i>E</i>. <i>coli</i> core metabolism.

<p>a) Calvin cycle, b) rTCA cycle, c) 3HP-4HB cycle, d) DC-4HB cycle, e) PyrS-PyrC-Glx bicycle, f) PyrS-PEPC-Glx bicycle. Reactions that are part of carbon fixation pathways (red arrows), reactions carrying flux (bold red arrows) and not carrying flux (thin arrows) in the FBA simulations are indicated. ATP (green), electron donors (blue) and CO₂/HCO₃^- (yellow) are color coded. Reactions of some non-branching pathways were lumped into one reaction for better visualization. For more detailed maps of the carbon fixation pathways see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157851#pone.0157851.g002" target="_blank">Fig 2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157851#pone.0157851.s001" target="_blank">S1</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157851#pone.0157851.s004" target="_blank">S4</a> Figs.</p

Main <i>in silico</i> results for all analyzed carbon fixation pathways.

<p>Results shown are based on a high flux of PPR proton pumping of 50 mmol/gCDW/h. We also included the number of enzymes natively present or already heterologously, functionally expressed <i>in vivo</i> in <i>E</i>. <i>coli</i> (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157851#pone.0157851.s021" target="_blank">S3 Text</a>).</p