Extended Data Fig. 3: In vitro activity of NphB and HlPT. | Nature

Extended Data Fig. 3: In vitro activity of NphB and HlPT.

From: Complete biosynthesis of cannabinoids and their unnatural analogues in yeast

Extended Data Fig. 3

All LC–MS chromatograms were selected for the theoretical m/z values of the respective compounds of interest (Extended Data Table 3). Chromatography gradient for cannabinoid analogues was used. Signals were compared to genuine standards. a, Purified NphB catalysed the condensation of GPP and olivetolic acid to CBGA when incubated with 5 mM olivetolic acid and 5 mM GPP at room temperature for 24 h. The enzyme produced at least one other isomer of CBGA, which is consistent with previous reports18. Boiling NphB (NphB ΔT) abolished activity. b, Microsomal fractions were prepared from yCAN21, yCAN22, and yCAN23 that expressed HlPT1L-T (1L-T), HlPT1L-T and HlPT2-T (1L-T/2-T), and HlPT2-T (2-T), respectively. Incubation with 5 mM olivetolic acid and 5 mM GPP for 24 h at 30 °C yielded CBGA as well as several isomers. CBGA production was not observed when incubating HlPT1L-T and HlPT2-T with their native substrates phlorisovalerophenone and dimethylallyl diphosphate (neg).

Back to article page