Fig. 6: The hydroxylase activity of three Physaria FAH12s.
From: Chromosome-level assembly and analysis of three hydroxy fatty acid-producing Physaria species
a The expression of the FAH12 genes across three different tissues of three Physaria species, alongside the proportion of lesquerolic acid to total seed fatty acids in their seeds. The left y-axis represents the expression levels (TPM) of the FAH12 genes in early developing seeds (EDS, yellow folded plot), middle developing seeds (MDS, red folded plot), and late developing seeds (LDS, blue folded plot). Meanwhile, the right y-axis indicates the proportion of lesquerolic acid relative to the total fatty acid content in seeds (gray folded plot). b Activity assay. Assay was performed for three Physaria FAH12s and PfFAH12 mutants in mammalian cells. We analyzed the content of product ricinoleic acid (C18:1-OH) using GC-FID. The data from three independent measurements are averaged, and the error indicates SD. Statistical analysis: unpaired two-tailed t-tests compared with PfFAH12WT. *p < 0.05, **p < 0.01 ***p < 0.001, ****p < 0.0001. n = 3 biological replicates. c Structural visualization of three Physaria FAH12 AlphaFold models in complex with oleic acid, respectively. The docked models are shown in surface and cartoon representation. The histidine motifs in active pocket and substrate are shown in stick representation. d Structural superimposition of these three Physaria FAH12 AlphaFold models. Two pairs of residues predicted to cause differences in the activity of FAH12 among three Physaria are zoomed-in and shown in stick representation. e The ricinoleic acid (C18:1-OH) content in T3 mature seeds. Values are means ± SD (n = 5 biological replicates). Different letters indicate differences at p < 0.05 using one-way ANOVA. Source data are provided as a Source Data file.
