Fig. 1: De novo alstonine and serpentine production in yeast.
From: Biosynthesis of natural and halogenated plant monoterpene indole alkaloids in yeast
a, Integration of plant biosynthetic pathways with native yeast metabolic pathways to produce alstonine and serpentine. IPP, isopentenyl pyrophosphate; DMAPP, dimethylallyl pyrophosphate; GPPS, GPP synthase; FPSN144W, FPP synthase N144W variant; CPR, NADPH-cytochrome P450 reductase; CYB5, cytochrome b5; GES, geraniol synthase; G8H, geraniol 8-hydroxylase; 8HGO, 8-hydroxygeraniol oxidoreductase; ISY, iridoid synthase; IO, iridoid oxidase; CYPADH, alcohol dehydrogenase 2; 7DLGT, 7-deoxyloganetic acid glucosyl transferase; 7DLH, 7-deoxyloganic acid hydroxylase; LAMT, loganic acid O-methyltransferase; TDC, tryptophan decarboxylase; SLS, secologanin synthase; STR, strictosidine synthase. b, Screen of AS candidates in YPD cultivation medium. Gene candidates are linked to strain identifiers as follows: RteAS1 (Sc87), RteAS2 (Sc88), RteAS3 (Sc90), RteAS4 (Sc92), RteAS5 (Sc94), CroAS_nat (Sc96), RseSBE_nat (Sc97), GseSBE_nat (Sc98), CroAS2_nat (Sc100), CroAS2 (Sc101), CroAS (Sc102), RseSBE (Sc103), GseSBE (Sc104) and CroSS_nat (Sc157) and a negative-control strain (Sc86). c,d, Representative production profiles for alstonine (c), serpentine (d) and pathway intermediates using a small-scale fed-batch process for strains Sc112 and Sc85, respectively, cultivated in 1 ml of 3× SC medium supplemented with 3 mM tryptophan. For b, n = 3, and error bars represent 1 s.d. from the mean, with data points overlaid as black dots.
