Engineering of BVMOs

Baeyer-Villiger monooxygenase (BVMOMtbs) Mycobacterium tuberculosis H37Rv 2006 [23] 2006 [23] [23, 24] 

Cydohexanone monooxygenase (CHMORhodoisa) Rhodococcus Phil and Phi2 2003 [30] 2003 [30] [26, 32[ 

Linear ketone monooxygenase (BVMOpaJ Pseudomonas fluorescens DSM 50106 2006 [42] 2006 [42] [42] 

Such long-distance mutations typically do not strongly influence the substrate acceptance profile of enzymes. In fact, it appears that, for effectively altering the substrate specificity of an enzyme, one should preferentially target active site residues. Such a targeted mutagenesis of first shell residues has been shown to result in dramatic changes in substrate specificity and/or enantioselectivity [49]. However, 

Future BVMO redesign studies can exploit the localization of these hotspots by simultaneously mutating the respective residues that determine the plasticity of the substrate-binding pocket of BVMOs. In this context it is noteworthy that, for several mutations in CHMO, an extension of substrate acceptance was observed with concomitant high turnover to allow for preparative exploitation [56]. By this strategy, novel valuable BVMOs can be created that extend the catalytic potential of the presently available BVMOs and may ultimately combine substrate promiscuity with other enzymatic properties, for example thermal stability in case of PAMO. 

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