Journal
ACS SYNTHETIC BIOLOGY
Volume 6, Issue 3, Pages 416-420Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.6b00297
Keywords
codon mutagenesis; directed evolution; halogenase; prolyl oligopeptidase; cytochrome P450
Categories
Funding
- NIH [1R01GM115665]
- U.S. Army Research Laboratory
- U.S. Army Research Office [62247-LS]
- NSF predoctoral fellowship [DGE-1144082]
- University of Chicago Department of Chemistry Helen Sellei-Beretvas Fellowship
- ARCS Scholar Award
Ask authors/readers for more resources
Directed evolution is a powerful tool for optimizing enzymes, and mutagenesis methods that improve enzyme library quality can significantly expedite the evolution process. Here, we report a simple method for targeted combinatorial codon mutagenesis (CCM). To demonstrate the utility of this method for protein engineering, CCM libraries were constructed for cytochrome P450(BM3), pfu prolyl oligopeptidase, and the flavin-dependent halogenase RebH; 10-26 sites were targeted for codon mutagenesis in each of these enzymes, and libraries with a tunable average of 1-7 codon mutations per gene were generated. Each of these libraries provided improved enzymes for their respective transformations, which highlights the generality, simplicity, and tunability of CCM for targeted protein engineering.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available