Extended Data Fig. 6. Screening of iridoid synthase (ISY) and cyclase (CYC) combinations for efficient conversion of 8-oxogeranial to cis-trans-nepetalactol.
Combinations of ISY and CYC were transformed into the MIA-BKV-1 strain (MIA-BJ without ISY/CYC). All ISYs and CYCs are under the control of GAL1 or GAL10 promoters. All yeast transformants (three biological replicates) were grown in SC medium (lacking histidine and leucine for selection of the two plasmids) containing 1.5% glucose and 1.5% galactose as carbon sources. All cultures were supplemented with 100 μM 8-oxogeranial and 100 μM tryptamine. After 3 days, additional 0.5% galactose and 100 μM 8-oxogeranial were added to the culture. Loganic acid, loganin, secologanin and strictosidine were measured and combined values were used as an approximate for total production of cis-trans-nepetalactol. EV, empty vector. Data presented are mean ± s.d. (n = 3). n.s. not significant; * p-value < 0.05; ** p-value < 0.01. Student’s two-tailed t-test. More statistical analysis is available in the source data file.