The amount of cholesterol made by many plants is not negligible. Whereas cholesterogenesis in animals was elucidated decades ago, the plant pathway has remained enigmatic. Among other roles, cholesterol is a key precursor for thousands of bioactive plant metabolites, including the well-known Solanum steroidal glycoalkaloids. Integrating tomato transcript and protein co-expression data revealed candidate genes putatively associated with cholesterol biosynthesis. A combination of functional assays including gene silencing, examination of recombinant enzyme activity and yeast mutant complementation suggests the cholesterol pathway comprises 12 enzymes acting in 10 steps. It appears that half of the cholesterogenesis-specific enzymes evolved through gene duplication and divergence from phytosterol biosynthetic enzymes, whereas others act reciprocally in both cholesterol and phytosterol metabolism. Our findings provide a unique example of nature's capacity to exploit existing protein folds and catalytic machineries from primary metabolism to assemble a new, multi-step metabolic pathway. Finally, the engineering of a ‘high-cholesterol’ model plant underscores the future value of our gene toolbox to produce high-value steroidal compounds via synthetic biology.
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We are grateful to D. Twafik for useful suggestions in phylogenetic analysis. A.A. is the incumbent of the Peter J. Cohn Professorial Chair. We thank the Adelis Foundation, the Leona M. and Harry B. Helmsley Charitable Trust, the Jeanne and Joseph Nissim Foundation for Life Sciences, Tom and Sondra Rykoff Family Foundation Research and the Raymond Burton Plant Genome Research Fund for supporting the laboratory activity of A.A. The work was supported by the Israel Science Foundation (ISF Grant No. 1805/15) and the European Research Council (ERC; SAMIT-FP7) personal grants to A.A. P.D.S. is grateful to the Planning and Budgeting Committee of the Council for Higher Education, Israel for the VATAT fellowship. The research in the laboratory of A.G. was financially supported by the VIB International PhD Fellowship Program (fellowship to P.A.) and the Research Foundation Flanders (postdoctoral fellowships to J.P. and L.P.). A.K was supported by a short-term EMBO fellowship (EMBO-ASTF-146-2014). The research in the laboratories of A.A. and A.G. was supported by the European Union Seventh Framework Program FP7/2007–2013 under grant agreement no. 613692–TriForC.
The authors declare no competing financial interests.
Supplementary Information. (PDF 32015 kb)
Detailed list of genes. (XLSX 18 kb)
Amino acid sequences. (PDF 286 kb)
Chemical structure: 4α-methyl-5α-cholest-7-en-3βol. (CDX 3 kb)
Chemical structure: 4α-methyl-ergostatrienol. (CDX 3 kb)
Chemical structure: 4α-methyl-24,25-dihydrozymosterol. (CDX 3 kb)
Chemical structure: 4α-methylcholesta-8,14-dien-3β-ol. (CDX 3 kb)
Chemical structure: 4α-methyl-ergostadienol. (CDX 3 kb)
Chemical structure: 7-dehydrocholesterol. (CDX 3 kb)
Chemical structure: 24-ethylidenelophenol. (CDX 3 kb)
Chemical structure: 24-methylenecholesterol. (CDX 3 kb)
Chemical structure: 24-methylenecycloartanol. (CDX 3 kb)
Chemical structure: 24-methylenelophenol. (CDX 3 kb)
Chemical structure: 31-nor-24,25-dihydrolanosterol. (CDX 3 kb)
Chemical structure: 31-norcycloartano. (CDX 3 kb)
Chemical structure: α-chaconine. (CDX 3 kb)
Chemical structure: α-solanine. (CDX 4 kb)
Chemical structure: α-tomatine. (CDX 5 kb)
Chemical structure: β-sitosterol. (CDX 3 kb)
Chemical structure: β-amyrin. (CDX 3 kb)
Chemical structure: campesterol. (CDX 3 kb)
Chemical structure: cholesta-7-en-3β-ol. (CDX 3 kb)
Chemical structure: cholesterol. (CDX 3 kb)
Chemical structure: cycloartanol. (CDX 3 kb)
Chemical structure: cycloartenol. (CDX 3 kb)
Chemical structure: cycloeucalenol. (CDX 3 kb)
Chemical structure ( Delta 5,7 avenasterol) (CDX 3 kb)
chemical structure(Delta 5, Episterol) (CDX 3 kb)
Chemical structure (structure 2): 4,4-dimethylcholesta-8,24-dien-3β-ol (CDX 3 kb)
Chemical structure: δ-7-avenasterol. (CDX 3 kb)
Chemical structure: episterol. (CDX 3 kb)
Chemical structure: esculeoside A. (CDX 7 kb)
Chemical structure: isofucosterol. (CDX 3 kb)
Chemical structure: lanosterol. (CDX 3 kb)
Chemical structure: obtusifoliol. (CDX 3 kb)
Chemical structure: stigmasterol. (CDX 3 kb)
Chemical structure (structure 1): 4,4-dimethylcholesta-8,14(15),24-trien-3β-ol. (CDX 3 kb)
Chemical structure (structure 3): cholesta-8,24-dien-3β-ol. (CDX 3 kb)
Chemical structure (structure 4): cholesta-7,24-dien-3β-ol. (CDX 3 kb)
Chemical structure (structure 5): 7-dehydrodesmosterol. (CDX 3 kb)
Chemical structure (structure 6): desmosterol. (CDX 3 kb)
Chemical structure: uttroside B. (CDX 7 kb)
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Sonawane, P., Pollier, J., Panda, S. et al. Plant cholesterol biosynthetic pathway overlaps with phytosterol metabolism. Nature Plants 3, 16205 (2017). https://doi.org/10.1038/nplants.2016.205
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