(A) Bacterialization of the ultimate yeast gene in the heme biosynthesis pathway results in a distinct pink colony phenotype on YPD agar medium. In contrast, wild type BY4741 strain colonies appear as creamy-white. (B) Acetate-extracted secreted products from the pink Sc-hem15Δ::Ec-hemH strains show strongly enhanced fluorescence at 635 nm (excitation 399 nm), comparable to a protoporphyrin IX standard and unlike a heme standard or extracts from the parental BY4741 strain. The introduction of an MLS to the bacterialized yeast strain (Sc-hem15Δ::Ec-MLS-hemH) significantly reduced protoporphyrin IX secretion, while deletion of the MLS from the native yeast locus in strain Sc-ΔMLS-HEM15 caused several strains to increase protoporphyrin IX secretion.
DOI:
http://dx.doi.org/10.7554/eLife.25093.014