A fluorogenic assay for transketolase from Saccharomyces cerevisiae

In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We examined the use of the fluorogenic compound I as donor substrate of TK, which is itself non fluorescent but releases a fluorescent product: umbelliferone. (C) 2003 Elsevier Science Ltd. All rights reserved.