Characterization of hybrid biphenyl dioxygenases obtained by recombining Burkholderia sp. strain LB400 bphA with the homologous gene of Comamonas testosteroni B-356

Can J Microbiol. 2001 Nov;47(11):1025-32.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Burkholderia / genetics*
  • Chimera
  • Comamonas testosteroni / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Iron-Sulfur Proteins*
  • Oxygenases / genetics*
  • Polychlorinated Biphenyls / metabolism*
  • Protein Engineering*

Substances

  • Iron-Sulfur Proteins
  • Polychlorinated Biphenyls
  • Oxygenases
  • biphenyl-2,3-dioxygenase