Directed Evolution and Genetic Engineering of EHs

Due to the complexity of biocatalyst engineering and its limitations when using site-directed mutagenesis of a few mutational sites, in vitro enzyme evolution has emerged as an additional powerful tool for improving enzymes [48]. Using this 

8 EPOXIDE HYDROLASES AND THEIR APPLICATION IN ORGANIC SYNTHESIS 

TABLE 8.1 Selected EH Variants Evolved toward Higher Enantioselectivities 

Two-step screening (1) growth on epoxide-containing agar plates and (2) MS-based system Chiral HPLC-based test 

Reetz and Zheng [50] NDT codon randomization use of chaperones Reetz et al. [51] NDT codon randomization 

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