Characterization of hybrid biphenyl dioxygenases obtained by recombining Burkholderia sp. strain LB400 bphA with the homologous gene of Comamonas testosteroni B-356.
The bacterial degradation of polychlorinated biphenyls depends on the ability of the enzyme biphenyl 2,3-dioxygenase (BPDO) to catalyze their oxygenation. Analysis of hybrid BPDOs obtained using common restriction sites to exchange large DNA fragments between LB400 bphA and B-356 bphA showed that the C-terminal portion of LB400 alpha subunit can withstand extensive structural modifications, and that these modifications can change the catalytic properties of the enzyme. On the other hand, exchanging the C-terminal portion of B-356 BPDO alpha subunit with that of LB400 alpha subunit generated inactive chimeras. Data encourage an enzyme engineering approach, consisting of introducing extensive modifications of the C-terminal portion of LB400 bphA to extend BPDO catalytic properties toward polychlorinated biphenyls.
Barriault
D
D
Institut national de la recherche scientifique, INRS-Institut Armand-Frappier, Centre de microbiologie et biotechnologie, Université du Québec, Pointe-Claire, Canada.
Simard
C
C
Chatel
H
H
Sylvestre
M
M
D019117:Burkholderia / Q000235:genetics*; D002678:Chimera; D020604:Comamonas testosteroni / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D007506:Iron-Sulfur Proteins; D010105:Oxygenases / Q000235:genetics*; D011078:Polychlorinated Biphenyls / Q000378:metabolism*; D015202:Protein Engineering
D007506:Iron-Sulfur Proteins; D011078:Polychlorinated Biphenyls; D010105:Oxygenases; C074161:biphenyl-2,3-dioxygenase