Fig. 5: Metabolic engineering in Nicotiana benthamiana for producing the key intermediate 24-methyldesmosterol (2). | Nature Communications

Fig. 5: Metabolic engineering in Nicotiana benthamiana for producing the key intermediate 24-methyldesmosterol (2).

From: Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis

Fig. 5

a Overview of the two different engineering strategies that were attempted in this work. b Effect of phytosterol pathway gene overexpression (strategy II) on production of 24-methyldesmosterol (2). Total ion current (TIC) GC-MS chromatograms of representative samples from transient expression in N. benthamiana. The bottom bar plot indicates relative 24-methyldesmosterol (2) levels when a single gene of the 15-gene set from tHMGR to 24ISO was left out. Bar plots show means ± SEM and data points of three biological replicates. * P < 0.05, ** P < 0.01, NS not significant by Student’s two-tailed unpaired t test; exact P values are indicated. LOD: limit of detection, LOQ, limit of quantification. c GC-MS TIC chromatograms showing formation of two shunt products 7 and 8 dependent on 24ISO in N. benthamiana. d Proposed formation of shunt products 7 and 8 by background epoxidation of the Δ24 double bond in N. benthamiana. Nicotiana_benthamiana icon by Connor-Tansley (https://github.com/Ctansley) is licensed under CC-BY 4.0 Unported (https://creativecommons.org/licenses/by/4.0/) and was used without modifications. Source data are provided as a Source Data file.

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