Engineering escherichia coli for production of functionalized terpenoids using plant p450s

ABSTRACT Terpenoids are a highly diverse class of natural products that have historically provided a rich source for discovery of pharmacologically active small molecules1, such as

paclitaxel (Taxol) and artemisinin. Unfortunately, these secondary metabolites are typically produced in low abundance in their host organism, and their isolation consequently suffers from

low yields and high consumption of natural resources. Furthermore, chemical synthesis of terpenoids can also be difficult to scale for industrial production. For these reasons, an attractive

alternative strategy is to engineer metabolic pathways for production of pharmaceuticals or their precursors in a microbial host such as _Escherichia coli_. A key step is developing methods

to carry out cytochrome P450 (P450)-based oxidation chemistry _in vivo_. Toward this goal, we have assembled two heterologous pathways for the biosynthesis of plant-derived terpenoid

natural products, and we present the first examples of _in vivo_ production of functionalized terpenoids in _E. coli_ at high titer using native plant P450s. Access through your institution

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OTHERS DESIGN OF A REDOX-PROFICIENT _ESCHERICHIA COLI_ FOR SCREENING TERPENOIDS AND MODIFYING CYTOCHROME P450S Article 16 November 2023 ENGINEERING THE AMOEBA _DICTYOSTELIUM DISCOIDEUM_ FOR

BIOSYNTHESIS OF A CANNABINOID PRECURSOR AND OTHER POLYKETIDES Article 06 January 2022 A COMPUTATIONAL WORKFLOW FOR THE EXPANSION OF HETEROLOGOUS BIOSYNTHETIC PATHWAYS TO NATURAL PRODUCT

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expression. _Science_ 307, 1317–1321 (2005). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We thank X.-Y. Chen (Shanghai Institutes for Biological Sciences) for the

native _CAS_ and _CAH_ genes, L. Anthony (Amyris Biotechnologies) for the pAM92 plasmid, K. Fisher (Amyris Biotechnologies) for authentic standards, P. Ortiz de Montellano (University of

California, San Francisco) for the pCWori plasmid and J. Minshull (DNA 2.0), Y. Yoshikuni, D. Pitera, S. Withers and E. Paradise for helpful discussions. M.C.Y. Chang acknowledges a

postdoctoral fellowship from the Jane Coffin Childs Memorial Fund. Funding was provided by the Institute for OneWorld Health through generous support by the Bill and Melinda Gates

Foundation. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * California Institute for Quantitative Biomedical Research, University of California, Berkeley, Berkeley, 94720-3224, California, USA

Michelle C Y Chang, Rachel A Eachus, William Trieu, Dae-Kyun Ro & Jay D Keasling * Department of Chemical Engineering, Department of Bioengineering, University of California, Berkeley,

Berkeley, 94720-1462, California, USA Jay D Keasling * Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, 94720-3224, California, USA Jay D Keasling Authors *

Michelle C Y Chang View author publications You can also search for this author inPubMed Google Scholar * Rachel A Eachus View author publications You can also search for this author

inPubMed Google Scholar * William Trieu View author publications You can also search for this author inPubMed Google Scholar * Dae-Kyun Ro View author publications You can also search for

this author inPubMed Google Scholar * Jay D Keasling View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS M.C.Y.C. carried out experiments with

assistance from R.A.E. and W.T. M.C.Y.C. designed the experiments with input from D.-K.R. M.C.Y.C. and J.D.K. wrote the manuscript. All authors discussed the results and commented on the

manuscript. CORRESPONDING AUTHOR Correspondence to Jay D Keasling. ETHICS DECLARATIONS COMPETING INTERESTS J.D.K. owns shares of Amyris Biotechnologies, a company that is currently using the

technology described here to produce the antimalarial drug artemisinin. Neither Amyris nor the University of California will make any profit (or royalties) from the sale of artemisinin in

the developing world. SUPPLEMENTARY INFORMATION SUPPLEMENTARY METHODS (PDF 7055 KB) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Chang, M., Eachus,

R., Trieu, W. _et al._ Engineering _Escherichia coli_ for production of functionalized terpenoids using plant P450s. _Nat Chem Biol_ 3, 274–277 (2007). https://doi.org/10.1038/nchembio875

Download citation * Received: 22 February 2007 * Accepted: 21 March 2007 * Published: 15 April 2007 * Issue Date: May 2007 * DOI: https://doi.org/10.1038/nchembio875 SHARE THIS ARTICLE

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