A high-throughput fluorescence-based glycosyltransferase screen and its application in directed evolution
- 14 February 2008
- journal article
- research article
- Published by Springer Nature in Nature Protocols
- Vol. 3 (3) , 357-362
- https://doi.org/10.1038/nprot.2007.538
Abstract
This protocol details the application of a high-throughput fluorescence-based screen, in conjunction with error-prone PCR/saturation mutagenesis, for altering the proficiency and/or promiscuity of a secondary metabolite glycosyltransferase (GT) via directed evolution. Given the structural and mechanistic similarities among secondary metabolite-associated GTs, this approach may provide a template for engineering other members of the GT-B superfamily.Keywords
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