The haem monooxygenase cytochrome P450cam has been engineered to oxidise the gaseous alkanes butane and propane to butan-2-ol and propan-2-ol, respectively, by the use of bulky amino acid substitutions to reduce the volume of the substrate pocket and thus improve the enzyme-substrate fit: the F87W/Y96F/T101L/V247L mutant oxidizes butane with a turnover rate of 750 min-1 and 95% yield based on NADH consumed while the wild-type enzyme has an activity of 0.4 min-1 with 4% yield.
Keywords:
Binding Sites
,Butanes
,Butanols
,Camphor 5-Monooxygenase
,Kinetics
,Mutagenesis, Site-Directed
,Oxidation-Reduction
,Propane
,Propanols
,Protein Engineering
,Pseudomonas putida
