Abstract
Microbial genome sequences are providing a wealth of information on new enzymes that have considerable potential as biocatalysts. The recently sequenced genome of Rhodococcus jostii RHA1, for example, has revealed an impressive array of catabolic enzymes, including many putative Baeyer-Villiger monooxygenases (BVMOs). We,have cloned 23 target BVMO sequences from the genome of R. jostii RHA1 and heterologously expressed 13 of these as soluble proteins to unearth new substrate specificities and selectivities. Whole-cell biocatalysts expressing the genes were screened against seven different test substrates. Each of these catalysts displayed activity toward at least three ketones. We observed a remarkable diversity of both regio- and enantioselectivity among the BVMOs from R. jostii RHA1 for the transformation of two chiral substrates, with some enzymes displaying high enantioselectivity for the isomers of 2-methylcyclopenta none. With the notable exception of the product of gene ro03437, named MO14, the biocatalysts' sequences correlated well with their respective activities and selectivities. This correlation allowed the identification of sequence motifs specific to subgroups of the BVMOs from R. jostii and other organisms. Overall, the data improve predictive models of BVMO activity from sequence and suggest new avenues to pursue in engineering these enzymes.
| Original language | English |
|---|---|
| Pages (from-to) | 1208-1217 |
| Number of pages | 10 |
| Journal | Chembiochem |
| Volume | 10 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 4 May 2009 |
Keywords
- Baeyer-Villiger reaction
- enzyme catalysis
- ketones
- oxidoreductases
- stereoselectivity
- XANTHOBACTER SP ZL5
- PHENYLACETONE MONOOXYGENASE
- MYCOBACTERIUM-TUBERCULOSIS
- BICYCLIC KETONES
- BIOCATALYSTS
- OXIDATION
- EXPRESSION
- CLONING
- ENANTIOSELECTIVITY
- PURIFICATION