A novel semi-biosynthetic route for artemisinin production using engineered substrate-promiscuous P450BM3

Jeffrey A. Dietrich, Yasuo Yoshikuni, Karl J. Fisher, Frank X. Woolard, Denise Ockey, Derek J. McPhee, Neil S. Renninger, Michelle C.Y. Chang, David Baker, Jay D. Keasling

Research output: Contribution to journalArticlepeer-review

170 Scopus citations

Abstract

Production of fine chemicals from heterologous pathways in microbial hosts is frequently hindered by insufficient knowledge of the native metabolic pathway and its cognate enzymes; often the pathway is unresolved, and the enzymes lack detailed characterization. An alternative paradigm to using native pathways is de novo pathway design using well-characterized, substrate-promiscuous enzymes. We demonstrate this concept using P450BM3 from Bacillus megaterium. Using a computer model, we illustrate how key P450BM3 active site mutations enable binding of the non-native substrate amorphadiene. Incorporating these mutations into P450BM3 enabled the selective oxidation of amorphadiene artemisinic-11S,12-epoxide, at titers of 250 mg L-1 in E. coli. We also demonstrate high-yielding, selective transformations to dihydroartemisinic acid, the immediate precursor to the high-value antimalarial drug artemisinin.

Original languageEnglish (US)
Pages (from-to)261-267
Number of pages7
JournalACS chemical biology
Volume4
Issue number4
DOIs
StatePublished - Apr 17 2009
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine

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