Development of screening methods for transketolase activity and substrate scope

A new GC-based method is developed for the screening of activity and stereoselectivity of a wide range of polyols as substrates of transketolase (TK). Compared to previous screening methods, it shows higher reproducibility, sensitivity and range of detection. In combination with HPLC screening, it can be used efficiently to test mutant libraries obtained by directed evolution methods.

Screening for activity, substrates (acceptor and donor) and selectivity (including enantioselectivity) is reported and a novel automated method to screen libraries of transketolase mutants is described.