B 12 -dependent dehydratases with improved reaction kinetics

Details B 12 -dependent dehydratases with improved reaction kinetics B 12 -dependent dehydratases with improved reaction kinetics

2003-12-16

2008-08-12

Prouty, Rebecca (Department: 1652)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

C435S232000, C435S440000

Reexamination Certificate

active

07410754

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.

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