Supplementary data for Model-driven engineering of Cutaneotrichosporon oleaginosus ATCC 20509 for improved microbial oil production

Supplementary data corresponding to manuscript named Model-driven engineering of Cutaneotrichosporon oleaginosus ATCC 20509 for improved microbial oil production. 

The Supplementary material document contains supplementary figures and tables. The content of the figures and tables are indicated below. 

Figure S1. Plasmid map of pUC57NAT containing pGpd, nourseothricin acyltransferase gene and tGpd.

Figure S2. Plasmid maps of overexpression targets containing TEF1α promoter, ATP-citrate lyase gene, TEF1α terminator, TPI1 promoter, Acetyl-CoA carboxylase gene, TPI1, YAT1 promoter, threonine synthase gene, YAT1 terminator and ENO1 promoter, hydroxymethylglutaryl-CoA synthase gene, ENO1 terminator.

Table S2. Nucleotide sequences of promoters, genes, and terminators from C. oleaginosus.

Figure S3. Calibration curve of glycerol for calculating the glycerol concentration of medium.

Figure S4. Volcano plots displaying differentially expressed genes and fold change (log2) in expression levels in WT, Δ9 and Δ12 strains at low lipid accumulation vs high lipid accumulation conditions.

Figure S5. Flux distribution graphs of selected reactions for overexpression in C. oleaginosus.

Figure S6. Colony PCR products were run on 1 % agarose gel. The colony PCR was performed for WT, ACL, ACC and TS transformants.

Table S4. qPCR outputs, CT: The threshold cycle.

Table S5. Fatty acid profile of C. oleaginosus grown at minimal medium with or without supplement (biotin, thiamine, threonine, serine, and aspartate) at 96h.

Table S6. Lipid content, dry cell weight, and lipid weight of WT, ACL, ACC, TS, and HMGS C. oleaginosus at various C/N ratio minimal medium.

Table S7. Fatty acid profile of WT, ACL, ACC, HMGS, and TS grown at C/N30, 120, 175, 200, and 300 minimal medium at 96h.

Figure S7. Quadratic regression analysis on lipid accumulation, biomass and lipid content of wild-type, ACL, ACC, and TS C. oleaginosus at C/N 30, 120, 175, 200, 300.

Table S8. Regression equations, statistics of regression equations for lipid content, biomass, and lipid content of wild-type, ACL, ACC, and TS.

Table S9. Calculated optimum C/N ratios and responses (lipid content, biomass, and total lipid) by using built regression models for wild-type, ACL, ACC, and TS.

Authors: 

Zeynep Efsun Duman-Özdamara,b,c, Mattijs K. Julsingc, Janine A.C. Verbokkemc, Emil Wolbertc, Vitor A.P. Martins dos Santosa,b,d, Jeroen Hugenholtze,f, Maria Suarez-Diezb*

aBioprocess Engineering, Wageningen University & Research, 6708 PB, Wageningen, the Netherlands

bLaboratory of Systems and Synthetic Biology, Wageningen University & Research,  6708 WE, Wageningen, the Netherlands

cWageningen Food & Biobased Research, Wageningen University & Research, 6708 WE, Wageningen, The Netherlands

dLifeGlimmer GmbH, Berlin, 12163, Germany

eFaculty of Science Swammerdam Institute for Life Sciences, University of Amsterdam, 1090 GE Amsterdam, The Netherlands

fNoPalm Ingredients  BV, 6709 PA Wageningen, The Netherlands