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Fig. 3 | Biotechnology for Biofuels

Fig. 3

From: Engineering the oleaginous yeast Yarrowia lipolytica to produce limonene from waste cooking oil

Fig. 3

Effects of single-gene overexpression of genes involved in the MVA pathway on d-limonene or l-limonene production. a Effects of single-gene overexpression of genes involved in the MVA pathway on d-limonene production. Ten genes including ACOAAT1, ACOAAT2, HMGS, HMGR, MK, PMK, PMVADO, IPPDI, GGPPS and FPPS involved in MVA pathway were overexpressed individually with d-limonene synthase. b Effects of single-gene overexpression of genes involved in the MVA pathway on l-limonene production. Ten genes including ACOAAT1, ACOAAT2, HMGS, HMGR, MK, PMK, PMVADO, IPPDI, GGPPS and FPPS involved in MVA pathway were overexpressed individually with l-limonene synthase. Bars represent limonene yields and lines represent gene expression improvements over controls. Titers of d-limonene or l-limonene were quantified after 5 days of cultivation and the RNAs of ten genes were extracted after 3 days of cultivation in shake flasks with 30 mL of liquid YPD medium. Glucose was used as the carbon source. The dLS- or lLS-integrated strain cultivated in parallel was used as controls, and the gene encoding the β-actin protein was used as an internal standard. Relative gene expression measured relative to β-actin and normalized to controls was calculated as 2−ΔΔCT. All values presented are the mean of three biological replicates ± standard deviation

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