| | |  | | | | Record from Web of Science® | | | | | | |
| Designed divergent evolution of enzyme function |
|
|
| Author(s): Yoshikuni Y, Ferrin TE, Keasling JD |
| Source: NATURE Volume: 440 Issue: 7087 Pages: 1078-1082 Published: APR 20 2006 |
| Times Cited: 86 References: 30 |
| Abstract: It is generally believed that proteins with promiscuous functions divergently evolved to acquire higher specificity and activity(1-5), and that this process was highly dependent on the ability of proteins to alter their functions with a small number of amino acid substitutions (plasticity)(6). The application of this theory of divergent molecular evolution to promiscuous enzymes may allow us to design enzymes with more specificity and higher activity. Many structural and biochemical analyses have identified the active or binding site residues important for functional plasticity (plasticity residues)(6-10). To understand how these residues contribute to molecular evolution, and thereby formulate a design methodology, plasticity residues were probed in the active site of the promiscuous sesquiterpene synthase gamma-humulene synthase(11,12). Identified plasticity residues were systematically recombined based on a mathematical model in order to construct novel terpene synthases, each catalysing the synthesis of one or a few very different sesquiterpenes. Here we present the construction of seven specific and active synthases that use different reaction pathways to produce the specific and very different products. Creation of these enzymes demonstrates the feasibility of exploiting the underlying evolvability of this scaffold, and provides evidence that rational approaches based on these ideas are useful for enzyme design. |
| Document Type: Article |
| Language: English |
| Reprint Address: Keasling, JD (reprint author), Univ Calif Berkeley, UCSF UCB Joint Grad Grp Bioengn, Berkeley, CA 94720 USA |
Addresses:
1. Univ Calif Berkeley, UCSF UCB Joint Grad Grp Bioengn, Berkeley, CA 94720 USA
2. Univ Calif Berkeley, Dept Chem Engn, Berkeley, CA 94720 USA
3. Univ Calif Berkeley, Calif Inst Quantitat Biomed Res QB3, Berkeley, CA 94720 USA
4. Lawrence Berkeley Lab, Phys Biosci Div, Synthet Biol Dept, Berkeley, CA 94710 USA
5. Univ Calif San Francisco, Dept Pharmaceut Chem & Biopharmaceut Sci, San Francisco, CA 94143 USA |
| Publisher: NATURE PUBLISHING GROUP, MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND |
| Subject Category: Multidisciplinary Sciences |
| IDS Number: 034DL |
| ISSN: 0028-0836 |
| DOI: 10.1038/nature04607 |
|
| | | | | | | | | Record from Web of Science® | | | | | | | | | |