Six enzymes from mayapple that complete the biosynthetic pathway to the etoposide aglycone

Science. 2015 Sep 11;349(6253):1224-8. doi: 10.1126/science.aac7202.

Abstract

Podophyllotoxin is the natural product precursor of the chemotherapeutic etoposide, yet only part of its biosynthetic pathway is known. We used transcriptome mining in Podophyllum hexandrum (mayapple) to identify biosynthetic genes in the podophyllotoxin pathway. We selected 29 candidate genes to combinatorially express in Nicotiana benthamiana (tobacco) and identified six pathway enzymes, including an oxoglutarate-dependent dioxygenase that closes the core cyclohexane ring of the aryltetralin scaffold. By coexpressing 10 genes in tobacco-these 6 plus 4 previously discovered-we reconstitute the pathway to (-)-4'-desmethylepipodophyllotoxin (the etoposide aglycone), a naturally occurring lignan that is the immediate precursor of etoposide and, unlike podophyllotoxin, a potent topoisomerase inhibitor. Our results enable production of the etoposide aglycone in tobacco and circumvent the need for cultivation of mayapple and semisynthetic epimerization and demethylation of podophyllotoxin.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Biosynthetic Pathways / genetics
  • Etoposide / metabolism*
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Plant
  • Genetic Engineering*
  • Methylation
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism*
  • Molecular Sequence Data
  • Podophyllotoxin / analogs & derivatives*
  • Podophyllotoxin / biosynthesis
  • Podophyllotoxin / metabolism*
  • Podophyllum peltatum / enzymology*
  • Podophyllum peltatum / genetics
  • Tobacco / genetics
  • Tobacco / metabolism*
  • Topoisomerase Inhibitors / metabolism*
  • Transcriptome

Substances

  • Topoisomerase Inhibitors
  • Etoposide
  • Mixed Function Oxygenases
  • Podophyllotoxin

Associated data

  • GENBANK/KT390155
  • GENBANK/KT390156
  • GENBANK/KT390157
  • GENBANK/KT390158
  • GENBANK/KT390159
  • GENBANK/KT390160
  • GENBANK/KT390161
  • GENBANK/KT390162
  • GENBANK/KT390163
  • GENBANK/KT390164
  • GENBANK/KT390165
  • GENBANK/KT390166
  • GENBANK/KT390167
  • GENBANK/KT390168
  • GENBANK/KT390169
  • GENBANK/KT390170
  • GENBANK/KT390171
  • GENBANK/KT390172
  • GENBANK/KT390173
  • GENBANK/KT390174
  • GENBANK/KT390175
  • GENBANK/KT390176
  • GENBANK/KT390177
  • GENBANK/KT390178
  • GENBANK/KT390179
  • GENBANK/KT390180
  • GENBANK/KT390181
  • GENBANK/KT390182
  • SRA/SRP061783