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Fig. 4 | Microbial Cell Factories

Fig. 4

From: Heterologous production of raspberry ketone in the wine yeast Saccharomyces cerevisiae via pathway engineering and synthetic enzyme fusion

Fig. 4

Full de novo biosynthesis of raspberry ketone. a Codon optimized genes encoding phenylalanine ammonia lyase from Rhodosporidium toruloides PAL (RtPAL) and cinnamate-4-hydroxylase from Arabidopsis thaliana (AtC4H) were integrated at the HO locus of S. cerevisiae as either two independent genes or as a single ORF fused by either a flexible (f) or rigid (r) amino acid linker. All strains also contained the P. crispum coumarate CoA ligase 2 and benzalacetone synthase from R. palmatum ORFs fused by a flexible linker (Pc4CL2-f-RpBAS), positioned adjacently in the HO locus. Levels of raspberry ketone were assessed following five days growth at 22 °C in air-lock flasks in synthetic grape juice medium and assessed for raspberry ketone production via LC/MS. b A strain containing the Rhodosporidium toruloides PAL (RtPAL) and cinnamate-4-hydroxylase from Arabidopsis thaliana (AtC4H) as separate ORFs in addition to the P. crispum coumarate CoA ligase 2 and benzalacetone synthase from R. palmatum ORFs fused by rigid linker (Pc4CL2-r-RpBAS) were fermented in either airlock flasks (anaerobic) or standard flasks (aerobic) in synthetic grape juice medium and assessed for raspberry ketone production via LC/MS

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