Additional file 1 of Novel evolved Yarrowia lipolytica strains for enhanced growth and lipid content under high concentrations of crude glycerol
Creators: name: Tsirigka, Asimina
name: Theodosiou, Eleni
name: Patsios, Sotiris I.
name: Tsoureki, Antiopi
name: Andreadelli, Aggeliki
name: Papa, Elisavet
name: Aggeli, Amalia
name: Karabelas, Anastasios J.
name: Makris, Antonios M.
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Biotechnology
subject: Evolutionary Biology
subject: Chemical Sciences not elsewhere classified
subject: Developmental Biology
subject: Marine Biology
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Computational Biology

description: Additional file 1: Table S1. Osmolality of synthetic medium (SM), supplemented with different concentrations of pure and crude glycerol. Figure S1. Monitoring of cell morphology during the cultivation of Yarrowia lipolytica MUCL 28,849 wild type (WTpar) and the evolved strain YLE469, in 25 mL synthetic medium, containing 15% v/v pure glycerol. Letters indicate cultivation timepoints, (a) 24 h, (b) 48 h, (c) 72 h and (d) 96 h. YLE469 strain was obtained after EMS mutagenesis and adaptive laboratory evolution through 520 generations of gradual increase of pure glycerol into the medium (9–20% v/v). Snow-flake morphology was observed in two biological duplicates. Figure S2. Cell morphology of the evolved strain YLE469, cultivated in 25 mL synthetic medium, containing 15% v/v pure glycerol, for 48 h. YLE469 strain was obtained after EMS mutagenesis and adaptive laboratory evolution through 520 generations of gradual increase of pure glycerol into the medium (9–20% v/v). Snow-flake morphology was observed in two biological duplicates. Table S2. Number of reads obtained after sequencing and quality filtering of Yarrowia lipolytica 28,849 and evolved strains (after ALE). For each strain, the total number of sequences obtained for the three biological replicates is presented. Figure S3. Dotplot of gene ontology (GO) enrichment analysis for YLE111 evolved Yarrowia lipolytica strain. GO terms from top to bottom display biological process, cellular component, and molecular function domains. Figure S4. %OD drop of Yarrowia lipolytica cells during lag growth phase, grown in 50 mL synthetic medium containing different crude glycerol concentrations (5, 7.5, 10, 15, 20% v/v) using as inoculum (a) preculture from YPG broth (b) preculture cultivated first in YPG broth and then in synthetic medium for preadaptation. The drop was observed immediately after the inoculation and its duration increased with increasing glycerol concentration. Figure S5. Heatmap showing the expression levels of all genes in the strains WTpar, YLE111, YLE155, YLE198, YLE469 and YLE471. Gene expression levels were normalized across all samples using z-score. Blue and red color indicates high and low expression levels, respectively. Sample (columns) and gene (rows) clustering was performed based on the respective euclidean distances. Figure S6. Correlation between dry biomass (g/L) and optical density (OD). Dry biomass was collected from a bioreactor fermentation at different cultivation time points, while OD values were obtained from a submerged OD sensor. Table S3. Estimated carbon balance of WTpar and YLE155, cultivated in 1.75 L synthetic medium, supplemented with 20% v/v crude glycerol for 48 h. Fermentations were conducted in triplicates (N = 3). Carbon (mol) of glycerol and citric acid was estimated based on their chemical formula and calculated mass. According to Celińska et al., the percentage of carbon in dry biomass was equal to 50% (g/g) [77]. Table S4. Fatty acid (FA) composition of the intracellular lipids of WTpar and YLE155, cultivated in 1.75 L synthetic medium, supplemented with 20% v/v crude glycerol for 48 h. Fermentations were conducted in triplicates (N = 3).