Additional file 1 of Identification of lipid synthesis genes in Schizochytrium sp. and their application in improving eicosapentaenoic acid synthesis in Yarrowia lipolytica
Creators: name: Jia, Yu-Lei
name: Zhang, Qing-Ming
name: Du, Fei
name: Yang, Wen-Qian
name: Zhang, Zi-Xu
name: Xu, Ying-Shuang
name: Ma, Wang
name: Sun, Xiao-Man
name: Huang, He
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Biological Sciences not elsewhere classified
subject: Information Systems not elsewhere classified
subject: Hematology
subject: Computational Biology

description: Additional file 1: Table S1. Genes used in this study. Table S2. Strains and plasmids used in this study. Table S3. Primers used in this study. Figure S1. Biosynthesis pathway of triglyceride in Schizochytrium sp. HX-308. FAS fatty acid synthase, PKS polyketide synthase, EPA eicosapentaenoic acid, DPA docosapentenoic acid, DHA docosahexaenoic acid, G3P glycerol-3-phosphate, GPAT glycerol-3-phosphate acyltransferase, LPA lysophosphatidate, LPAAT lysophosphatidate acyltransferase, PA phosphatidate, PAP phosphatidic acid phosphatase, DG diacylglycerol, DGAT diacylglycerol acyltransferase, TG triacylglycerol. Figure S2. Amino acid sequence analysis of four DGATs. a Phylogenetic analysis of amino acid sequences of ScDGAT2A, ScDGAT2B, ScDGAT2C, and ScDGAT3. The neighbor-joining method was used to reconstruct the cladogram under the software MEGA 7. The scale bar 0.2 represents 20% divergence. The bracket after the species name represents the GenBank ID. b Protein sequence alignment of ScDGAT2A, ScDGAT2B, and ScDGAT2C with DGAT2s from five organisms. Figure S3. Predicated transmembrane domains for DGATs by TMHMM. Figure S4. Predicated subcellular localization of proteins for DGATs. Figure S5. Conserved domains detected in DGATs by NCBI Conserved Domains Search. Figure S6. PCR validation of DGAT genes integration into CK strain (a) and yl-EPA strain (b) genome. Figure S7. Gas chromatography (GC) analysis of ScDGAT2C, ScDGAT2AM, and ScDGAT2BM expressed sample. Figure S8. Effect of DGATs expression on fatty acids titer. Three biological replicates were used and mean values ± SD (n=3) are shown. Two-way ANOVA with Tukey’s multiple comparisons test. *P &lt; 0.05, **P &lt; 0.01, and ***p &lt; 0.001 compared with the yl-EPA-1. Figure S9. Effect of ScDGAT2C expression on PUFAs (a) and SFAs (b) composition in yl-EPA strain. Figure S10. Effect of ScDGAT2C expression on fatty acid composition in TG. Effects of ScDGAT2C on TG associated with C18:2 (a), 18:3 (b), and 20:4 (c) in yl-EPA strain. Three biological replicates were used and mean values ± SD (n = 3) are shown. Student’s t test was used for statistical analysis, and statistical significance is indicated as *P &lt; 0.05, **P &lt; 0.01, and ***p &lt; 0.001. Figure S11. Protein sequence alignment of ScGPAT1 and ScGPAT2 with GPATs from five organisms. Figure S12. Comparison of fatty acid composition between yl-EPA-1 and yl-EPA-8 strains. TFA total fatty acids. Three biological replicates were used and mean values ± SD (n = 3) are shown. Student’s t test was used for statistical analysis, and statistical significance is indicated as *P &lt; 0.05, **P &lt; 0.01, and ***p &lt; 0.001.