B12-dependent dehydratases with improved reaction kinetics
AssigneeE. I. Du Pont de Nemours and Company
InventorsKatharine J. Gibson, Der-Ing Liao, Xiao-Song Tang
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Abstract
Sequences of B12-dependent dehydratases with improved reaction kinetics are presented. Use of these B12-dependent dehydratases reduce the rate of the enzyme's suicide inactivation in the presence of glycerol and 1,3-propanediol. The enzymes were created using error-prone PCR and oligonucleotide-directed mutagenesis to target the DhaB1 gene, which encodes the α-subunit of glycerol dehydratase. Mutants with improved reaction kinetics were rapidly identified using high throughput assays.
Claims2
A nucleic acid sequence encoding a B12-dependent mutant dehydratase comprising an α-β subunit fusion and a γ-subunit having the polypeptide sequence set forth in SEQ ID NO:348, said α subunit having the polypeptide sequence set forth in SEQ ID NO:347 and said β subunit having the polypeptide sequence set forth in SEQ ID NO:3.
Dependencies are inferred from the claim text.