Additional file 1 of Reactor microbiome enriches vegetable oil with n-caproate and n-caprylate for potential functionalized feed additive production via extractive lactate-based chain elongation
Creators: name: Contreras-Dávila, Carlos A.
name: Zuidema, Norwin
name: Buisman, Cees J. N.
name: Strik, David P. B. T. B.
Publisher: name: figshare
Year: 2021
Subjects: subject: Biophysics
subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics
subject: Molecular Biology
subject: Biotechnology
subject: 59999 Environmental Sciences not elsewhere classified
subject: FOS: Earth and related environmental sciences
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Ecology
subject: 69999 Biological Sciences not elsewhere classified
subject: Inorganic Chemistry

description: Additional file 1: Figure S1. Metabolites concentration in the aqueous phase during extractive batch fermentation with: (A, D) sunflower oil and (B, E) oleyl alcohol. (C, F) Non-extractive fermentation singletons. Substrates were lactate (A–C) and food waste (D–F). Error bars depict duplicates absolute deviation from the average. Figure S2. Carboxylates concentration in solvents during batch extractive fermentation. Error bars depict duplicates absolute deviation from the average. Figure S3. Conversion rates for lactate-based chain elongation as measured over time in: (A) R1 and (B) R2. Batch periods correspond to days 46.8–54 in R1; days 47.8–55 and 57–64 in R2. Triangles show DNA sampling days (blue – suspended biomass samples, red – both suspended biomass and biofilm samples). DNA sample from period II-a was taken only for R1. Figure S4. Aqueous phase metabolites concentrations and pH over time in: (A) R1 and (B) R2. Figure S5. Selective extraction of n-caproate with sunflower oil from continuously fed synthetic medium. n-Caproate concentrations in sunflower oil as measured from back-extracted samples in: (A) R3 and (C) R4. Bubble size depicts extraction flux into the solvent based on cumulative carboxylates concentrations between two contiguous sampling points. Sankey diagrams show the carbon flux (mmol C d-1) for (B) R3 and (D) R4. The synthetic effluent contained lactate, acetate, propionate, n-butyrate and n-caproate fed to an HRT of 2 days and aqueous phase pH was controlled at 5.0. Nitrogen sources were left out to avoid microbial growth. Figure S6. Carbon flux (mmol C d-1) during non-extractive and extractive lactate-based chain elongation in (A–C) R1 and (D–F) R2. Sankey diagrams were built using data of periods I-a to I-b (non-extractive pH 5), period II-a (extractive pH 5.0) and period III (extractive pH 7.0). The unidentified missing carbon was assumed to be assimilated into biomass. Figure S7. Principal Components Analysis (PCA) plot of carboxylic acids composition of initial sunflower oil and MCC-enriched sunflower oil at different pH conditions. MCC-enriched sunflower oil samples were taken from both reactors at different times during extractive fermentation with sampling points same as in Figure 3. Carboxylic acids compositions was measured according to the ISO 15885 standard. Variables were scaled and centered for analysis. Top 15 variables are shown with vectors colored according to their contribution to variance in the PCA plot. Concentration ellipses depict confidence intervals with α = 0.05 for groups with n&gt;3. Principal Components Analysis (PCA) was done in R Studio using the prcomp function and visualized using the factoextra package [3]. Figure S8. (A) CSTR schematic and (B) picture. 1 – feed tank; 2 – oil sampling port; 3 – polyurethane foam; 4 – redox sensor; 5 – pH sensor; 6 – liquid level sensor; 7 – controller (Biocontroller ADI 1010, Applikon); 8 – stirring engine; 9 – solvent; 10 – fermentation broth; 11 – gas condenser (4°C); 12 – gas sampling port; 13 – gas meter (µFlow, Bioprocess Control); 14 – liquid sampling port; 15 – water jacket; 16 – effluent tank; 17 – acid tank (A). Table S1. Net production of carboxylates in extractive fermentation of lactate and food waste after 20 days. Table S2. Overview of chain elongation reactor R1 performance under non-extractive (I-a to I-c) and extractive (II-a to III) conditions. Table S3. Overview of chain elongation reactor R2 performance under non-extractive (I-a to I-c) and extractive (II-a to III) conditions. Table S4. Selected 16S rRNA gene amplicon sequence variants (ASV). Table S5. Stoichiometry of lactate-based chain elongation in R1 at pH 5.0 with(out) extraction with sunflower oil. Table S6. Overview of continuous chain elongation operational parameters.