Campus Units
Biochemistry, Biophysics and Molecular Biology, Roy J. Carver Department of
Document Type
Article
Publication Version
Accepted Manuscript
Publication Date
3-1-2016
Journal or Book Title
Proceedings of the National Academy of Sciences
Volume
113
Issue
9
First Page
2526
Last Page
2531
DOI
10.1073/pnas.1512096113
Abstract
The substrate specificity of enzymes from natural products’ metabolism is a topic of considerable interest, with potential biotechnological use implicit in the discovery of promiscuous enzymes. However, such studies are often limited by the availability of substrates and authentic standards for identification of the resulting products. Here, a modular metabolic engineering system is used in a combinatorial biosynthetic approach toward alleviating this restriction. In particular, for studies of the multiply reactive cytochrome P450, ent-kaurene oxidase (KO), which is involved in production of the diterpenoid plant hormone gibberellin. Many, but not all, plants make a variety of related diterpenes, whose structural similarity to ent-kaurene makes them potential substrates for KO. Use of combinatorial biosynthesis enabled analysis of more than 20 such potential substrates, as well as structural characterization of 12 resulting unknown products, providing some insight into the underlying structure–function relationships. These results highlight the utility of this approach for investigating the substrate specificity of enzymes from complex natural products’ biosynthesis.
Copyright Owner
The Author(s)
Copyright Date
2016
Language
en
File Format
application/pdf
Recommended Citation
Mafu, Sibongile; Jia, Meirong; Zi, Jiachen; Morrone, Dana; Wu, Yisheng; Xu, Meimei; Hillwig, Matthew L.; and Peters, Reuben J., "Probing the promiscuity of ent-kaurene oxidases via combinatorial biosynthesis" (2016). Biochemistry, Biophysics and Molecular Biology Publications. 289.
https://lib.dr.iastate.edu/bbmb_ag_pubs/289
Included in
Biotechnology Commons, Enzymes and Coenzymes Commons, Molecular Biology Commons, Natural Products Chemistry and Pharmacognosy Commons
Comments
This is a manuscript of an article published as Mafu, Sibongile, Meirong Jia, Jiachen Zi, Dana Morrone, Yisheng Wu, Meimei Xu, Matthew L. Hillwig, and Reuben J. Peters. "Probing the promiscuity of ent-kaurene oxidases via combinatorial biosynthesis." Proceedings of the National Academy of Sciences 113, no. 9 (2016): 2526-2531. DOI: 10.1073/pnas.1512096113. Posted with permission.