Engineering and improvement of the efficiency of a chimeric [P450cam-RhFRed reductase domain] enzyme.
A chimeric oxygenase, in which the P450cam domain was fused to the reductase host domains of a P450RhF from Rhodococcus sp. strain NCIMB 9784 was optimised to allow for a biotransformation at 30 mM substrate in 80% overall yield, with the linker region between P450 and FMN domain proving to be important for the effective biotransformation of (+)-camphor to 5-exo-hydroxycamphor.
Robin
Aélig
A
Manchester Interdisciplinary Biocentre, University of Manchester, UK.
Roberts
Gareth A
GA
Kisch
Johannes
J
Sabbadin
Federico
F
Grogan
Gideon
G
Bruce
Neil
N
Turner
Nicholas J
NJ
Flitsch
Sabine L
SL
D019475:Camphor 5-Monooxygenase / Q000737:chemistry* / Q000235:genetics*; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics*; D012240:Rhodococcus / Q000201:enzymology; D014170:Transformation, Genetic
D011993:Recombinant Fusion Proteins; D010088:Oxidoreductases; D019475:Camphor 5-Monooxygenase