Redesign of BVMOs

As mentioned above, the first BVMOs were already purified several decades ago. Subsequent biochemical studies have revealed that these enzymes are typically soluble and often easy to express at high levels in, e.g., E. coli. These features suggest that BVMOs are perfect candidates for X-ray crystallography studies. However, crystallization of several type I BVMOs has been attempted and proven 

While wild-type PAMO was unable to convert 2-phenylcyclohexanone efficiently, all deletion mutants readily accepted this ketone as substrate. All mutants also displayed a similar thermostability when compared with the parent enzyme. The most active mutant (deletion of S441 and A442) was used for examining its enantioselective properties. It was found that the mutant preferably formed the (/ )-enantiomer of the corresponding lactone E = 100). While CHMO also shows a similar enantioselective behavior, this PAMO deletion mutant is a better candidate for future applications due to its superior stability. This clearly demonstrates that PAMO can be used as parent enzyme to design thermostable BVMO variants. It also illustrates that the available crystal structure of PAMO will be of great help for BVMO redesign efforts.  

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