| abstract |
Sequences of B12-dependent dehydratases with improved reaction kinetics are presented, thus reducing the rate of the enzyme's suicide inactivation in the presence of glycerol and 1,3-propanediol. These enzymes were created using error-prone PCR and oligonucleotide-directed mutagenesis to target the DhaB1 gene, encoding the a-subunit of glycerol dehydratase. Mutants with improved reaction kinetics were rapidly identified using high throughput assays, also disclosed herein. |