Figure 5: Heterologous expression of Ban-Δ11, B. anynana pheromone production Δ11-desaturase gene.
From: Sex pheromone biosynthetic pathways are conserved between moths and the butterfly Bicyclus anynana
The graphs in (a) represent gas chromatogram traces of FAME from methanolysed InvSc1 S. cerevisiae yeast transformed with pYEX-CHT-Ban-Δ11 constructs without Cu2SO4 (upper panel, 0 mM Cu2+-control) and with addition of 2 mM Cu2SO4 (lower panel). In controls without promoter activation, a minor amount of Z11-16:acid (here and after, in the form of the corresponding methyl ester) is present through chain elongation from Z9-14:acid in yeast, a known activity inherent to the InvSc1 wild-type yeast strain. (b) In pYEX-CHT-Ban-Δ11 Cu2+-induced yeast samples, there is a threefold increase in the abundance of Z11-16:acid catalyzed by unsaturation of palmitic acid. Abundance ratios (±s.e.m.) of Z11-16:acid, 15:acid and Z9-15:acid in yeast samples bearing pYEX-Ban-Δ11 without (n=6) and with Cu2+ (n=4) relative to the IS (triheptadecenoin; converted into Z10-17:Me during base methanolysis). The y axes represent relative abundance. Independent sample t-tests were performed in IBM SPSS v.20 to determine whether addition of Cu2+ impacted on compound production (Z11-16:acid, ***P<0.001; 15:acid, P=0.729 (NS); Z9-15:acid, P=0.242 (NS); relative ratio between 15:acid and Z9-15:acid, P=0.348 (NS)). (c) Total amounts (± s.e.m.) of Z11-16:Me produced by yeast cell cultures bearing pYEX-Ban-Δ11 without (n=6; 1,948 ng±95) and with Cu2+ (n=4; 5,881 ng±355) (IBM SPSS v.20, independent samples t-test, ***P<0.001).
