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Figure 1 | Microbial Cell Factories

Figure 1

From: Combined metabolic engineering of precursor and co-factor supply to increase α-santalene production by Saccharomyces cerevisiae

Figure 1

Genetic engineering approach for increasing α-santalene production. (A) Expression plasmid pISP15 containing tHMG1 encoding truncated HMG-CoA reductase, a codon optimized santalene synthase gene (SanSyn opt ) P TEF1 and P PGK1 promoters as well as T ADH1 and T CYC1 terminator sequences. (B) Integrated cassettes, rectangles containing arrows represent the promoters and their directionality, pentagons the genes and empty squares the terminators. (C) Scheme of the engineered mevalonate, prenyl phosphate and ammonium assimilation pathways and FPP branch point; overexpressed and deleted genes are highlighted. Pathway intermediates: G6P: glucose-6-phosphate, Acetyl-CoAcyt: cytosolic acetyl-CoA, HMG-CoA: 3-hydroxy-3-methylglutaryl-CoA, MVA: mevalonate, MVA-P: phosphomevalonate, MVA-PP: diphosphomevalonate, IPP: isopentenyl diphosphate, DMAPP: dimethylallyl diphosphate, GPP: geranyl diphosphate, FPP: farnesyl diphosphate, FOH: farnesol. Overexpressed genes are tHMG1 (encoding truncated HMG-CoA reductase), ERG20 (encoding FPP synthase), GDH2 (encoding NAD-dependent glutamate dehydrogenase), and SanSyn opt (encoding α-santalene synthase). Deleted genes are GDH1 (encoding NADP-dependent glutamate dehydrogenase), LPP1 and DPP1 (both encoding lipid phosphate phosphatases). The promoter of the ERG9 gene (encoding squalene synthase) is replaced with P HXT1 . Genes whose promoters contain Upc2 binding sites are indicated with a grey arrow: ERG13 (encoding HMG-CoA synthase), ERG12 (encoding mevalonate kinase), and ERG8 (encoding phosphomevalonate kinase). Additional genes indicated are ERG10 (encoding acetoacetyl-CoA thiolase), ERG19 (encoding diphosphomevalonate decarboxylase) and IDI (encoding IPP isomerase).

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