A Novel Semi-biosynthetic Route for Artemisinin Production Using Engineered Substrate-Promiscuous P450BM3 [electronic resource].
- Berkeley, Calif. : Lawrence Berkeley National Laboratory, 2009. and Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy.
- Physical Description:
- 7 : digital, PDF file
- Additional Creators:
- Lawrence Berkeley National Laboratory and United States. Department of Energy. Office of Scientific and Technical Information
- Production of fine heterologus 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 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 activ site mutations enable binding of non-native substrate amorphadiene, incorporating these mutations into P450BM3 enabled the selective oxidation of amorphadiene arteminsinic-11s,12-epoxide, at titers of 250 mg L"1 in E. coli. We also demonstrate high-yeilding, selective transformations to dihydroartemisinic acid, the immediate precursor to the high value anti-malarial drug artemisinin.
- Published through SciTech Connect., 11/30/2009., "lbnl-2938e", American Chemical Society Journal of Chemical Biology 4 4 FT, and Fisher, Karl; Baker, David; Keasling, Jay; Renninger, Neil; Dietrich, Jeffrey; Yoshikuni, Yasuo; Woolard, Frank; Ockey, Denise; McPhee, Derek; Chang, Michelle.
- Funding Information:
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