Discovery of novel BVMOs

While the number of available recombinant BVMOs has grown significantly over the last few years, there is still a demand for other BVMOs to expand the biocat-alytic diversity. Most BVMOs that have been described are dedicated to efficiently

Figure 3: Illustration of overlapping substrate specificities of four BVMOs. Cyclohexanone monooxygenase (CHMO), cyclopentanone monooxygenase (CPMO), 4-hydroxyacetophenone monooxygenase (HAPMO) and phenylacetone monooxygenase (PAMO). For each enzyme, several typical substrates are shown.

Figure 3: Illustration of overlapping substrate specificities of four BVMOs. Cyclohexanone monooxygenase (CHMO), cyclopentanone monooxygenase (CPMO), 4-hydroxyacetophenone monooxygenase (HAPMO) and phenylacetone monooxygenase (PAMO). For each enzyme, several typical substrates are shown.

convert cyclohexanone and related cyclic aliphatic ketones. To cover a broader range of substrate types and enantio- and/or regioselectivities, new BVMOs have to be discovered. This can be done in a number of ways. In the past, it was common practice to isolate new microbes that were able to grow on a target substrate after which the respective enzyme/gene was retrieved. This approach has been successful in obtaining most of the presently available BVMOs (Table 1). Only PAMO and ethionamide monooxygenase have been discovered via other methods.1436 However, the classical approach of isolating a specific microorganism, enzyme purification and subsequently cloning is laborious, time consuming and often unsuccessful. Frequently, the enzyme responsible for the observed reaction is difficult to purify and hence the respective gene cannot be retrieved. Therefore, it is attractive to exploit other newly developed methods that circumvent these pitfalls.

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