Fig. 3: Coordinated action of BGC-localized GbCYPs (GbCYP7005C1, GbCYP7005C3, GbCYP867E38, and GbCYP867K1) advances ginkgolide biosynthesis. | Nature Communications

Fig. 3: Coordinated action of BGC-localized GbCYPs (GbCYP7005C1, GbCYP7005C3, GbCYP867E38, and GbCYP867K1) advances ginkgolide biosynthesis.

From: A gene cluster in Ginkgo biloba encodes unique multifunctional cytochrome P450s that initiate ginkgolide biosynthesis

Fig. 3

a LC–HRMS analysis of tobacco leaf extracts expressing GbLPS and CYP combinations as shown above each chromatogram. In all tobacco samples, CfDXS (C. forskohlii 1-deoxy-D-xylulose 5-phosphate synthase), CfGGPPS (C. forskohlii geranylgeranyl pyrophosphate synthase), and the P19 silencing suppressor are co-expressed. The chromatograms shown are extracted ion chromatograms (EICs, positive mode) for the m/z values of 187.1482; 203.1425; 317.2118; and 335.2218, each with a window of ±0.05. b LC–HRMS analysis of yeast terpenoid extracts from strains expressing SpGGPPS7, SctHMGR, GbLPS, GbPOR2, and CYP candidate combinations as shown above each chromatogram. The chromatograms shown are EICs (positive mode) for the m/z values of 187.1482; 203.1425; 638.2751; and 640.2890, each with a window of ±0.05. c proposed biosynthetic steps from ginkgosinoic acid A to ginkgosinoic acid C derivatives, and the responsible CYPs involved. The chemical structures of the compounds shown here (11, 13d, and 17) were elucidated using NMR spectroscopy. The putative intermediate ginkgosinoic acid C was not detected in its free form but was detected as glucoside (13d) in tobacco, or as glutathione conjugate (14–16) in yeast. Enzymatic in vitro removal of the glucose moiety from 13d gave rise to 17, a 1,4 benzoquinone derivative of ginkgosinoic acid C (Supplementary Fig. 9a). The numbering of carbon atoms follows the standard numbering of levopimaradiene. The Nicotiana benthamiana leaf image has been created by BioRender.com (2021).

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