|
|
|
|||||||
|
|||||||||
J Biol Chem, Vol. 275, Issue 17, 12430-12437, April 28, 2000
From the The oxygenase component of biphenyl dioxygenase
(BPDO) from Comamonas testosteroni B-356 dihydroxylates
biphenyl and some polychlorinated biphenyls (PCBs), thereby initiating
their degradation. Overexpressed, anaerobically purified BPDO had a
specific activity of 4.9 units/mg, and its oxygenase component appeared
to contain a full complement of Fe2S2 center
and catalytic iron. Oxygenase crystals in space group R3
were obtained under anaerobic conditions using polyethylene glycol as
the precipitant. X-ray diffraction was measured to 1.6 Å. Steady-state
kinetics assays demonstrated that BPDO had an apparent
kcat/Km for biphenyl of
(1.2 ± 0.1) × 106 M
Steady-state Kinetic Characterization and Crystallization of a
Polychlorinated Biphenyl-transforming Dioxygenase*
§,¶,
,, and
Department of Chemistry and Biochemistry,
Concordia University, Montreal, Quebec H3G 1M8, Canada, the
Department of Biological Sciences, Purdue University, West
Lafayette, Indiana 47907-1392, and the ** Department of Biochemistry,
Université Laval, Quebec City, Quebec G1K 7P4, Canada
1
s1 in air-saturated buffer. Moreover, BPDO transformed
dichlorobiphenyls (diClBs) in the following order of apparent
specificities: 3,3'- > 2,2'- > 4,4'-diClB. Strikingly, the ability of
BPDO to utilize O2 depended strongly on the biphenyl
substrate:
kcat/Km(O2) = (3.6 ± 0.3), (0.06 ± 0.02), and (0.4 ± 0.07) × 105 M1 s1 in the
presence of biphenyl and 2,2'- and 3,3'-diClBs, respectively. Moreover,
biphenyl/O2 consumed was 0.97, 0.44, 0.63, and 0.48 in the
presence of biphenyl and 2,2'-, 3,3'-, and 4,4'-diClBs, respectively.
Within experimental error, the balance of consumed O2 was
detected as H2O2. Thus, PCB congeners such as
2,2'-diClB exact a high energetic cost, produce a cytotoxic compound
(H2O2), and can inhibit degradation of other
congeners. Each of these effects would be predicted to inhibit the
aerobic microbial catabolism of PCBs.
*
This work was supported in part by Natural Sciences and
Engineering Research Council of Canada Strategic Grant STP0193182 (to
L. D. E. and J. B. P.) and by National Institutes of Health Grant
GM-52381 (to J. T. B.). Use of the Advanced Photon Source was
supported by the United States Department of Energy, Basic Energy
Sciences, Office of Energy Research, under Contract W-31-109-Eng-38. BioCARS Sector 14 was supported by National Institutes of Health National Center for Research Resources Grant RR-07707.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Microbiology and Immunology, University of British Columbia, 300-6174 University Blvd., Vancouver, BC V6T 1Z3, Canada. Tel.: 604-822-0042; Fax: 604-822-6041; E-mail: leltis@interchange.ubc.ca.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. K. Capyk, I. D'Angelo, N. C. Strynadka, and L. D. Eltis Characterization of 3-Ketosteroid 9{alpha}-Hydroxylase, a Rieske Oxygenase in the Cholesterol Degradation Pathway of Mycobacterium tuberculosis J. Biol. Chem., April 10, 2009; 284(15): 9937 - 9946. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. R. Master, N. Y. R. Agar, L. Gomez-Gil, J. B. Powlowski, W. W. Mohn, and L. D. Eltis Biphenyl Dioxygenase from an Arctic Isolate Is Not Cold Adapted Appl. Envir. Microbiol., June 15, 2008; 74(12): 3908 - 3911. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Patrauchan, C. Florizone, S. Eapen, L. Gomez-Gil, B. Sethuraman, M. Fukuda, J. Davies, W. W. Mohn, and L. D. Eltis Roles of Ring-Hydroxylating Dioxygenases in Styrene and Benzene Catabolism in Rhodococcus jostii RHA1 J. Bacteriol., January 1, 2008; 190(1): 37 - 47. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gomez-Gil, P. Kumar, D. Barriault, J. T. Bolin, M. Sylvestre, and L. D. Eltis Characterization of Biphenyl Dioxygenase of Pandoraea pnomenusa B-356 As a Potent Polychlorinated Biphenyl-Degrading Enzyme J. Bacteriol., August 1, 2007; 189(15): 5705 - 5715. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Camara, M. Seeger, M. Gonzalez, C. Standfuss-Gabisch, S. Kahl, and B. Hofer Generation by a Widely Applicable Approach of a Hybrid Dioxygenase Showing Improved Oxidation of Polychlorobiphenyls Appl. Envir. Microbiol., April 15, 2007; 73(8): 2682 - 2689. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Vezina, D. Barriault, and M. Sylvestre Family Shuffling of Soil DNA To Change the Regiospecificity of Burkholderia xenovorans LB400 Biphenyl Dioxygenase J. Bacteriol., February 1, 2007; 189(3): 779 - 788. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Parnell, J. Park, V. Denef, T. Tsoi, S. Hashsham, J. Quensen III, and J. M. Tiedje Coping with Polychlorinated Biphenyl (PCB) Toxicity: Physiological and Genome-Wide Responses of Burkholderia xenovorans LB400 to PCB-Mediated Stress Appl. Envir. Microbiol., October 1, 2006; 72(10): 6607 - 6614. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. R. Goncalves, H. Hara, D. Miyazawa, J. E. Davies, L. D. Eltis, and W. W. Mohn Transcriptomic Assessment of Isozymes in the Biphenyl Pathway of Rhodococcus sp. Strain RHA1 Appl. Envir. Microbiol., September 1, 2006; 72(9): 6183 - 6193. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Jouanneau and C. Meyer Purification and Characterization of an Arene cis-Dihydrodiol Dehydrogenase Endowed with Broad Substrate Specificity toward Polycyclic Aromatic Hydrocarbon Dihydrodiols. Appl. Envir. Microbiol., July 1, 2006; 72(7): 4726 - 4734. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Barriault and M. Sylvestre Evolution of the Biphenyl Dioxygenase BphA from Burkholderia xenovorans LB400 by Random Mutagenesis of Multiple Sites in Region III J. Biol. Chem., November 12, 2004; 279(46): 47480 - 47488. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Furukawa, H. Suenaga, and M. Goto Biphenyl Dioxygenases: Functional Versatilities and Directed Evolution J. Bacteriol., August 15, 2004; 186(16): 5189 - 5196. [Full Text] [PDF]
|
|
|
|
|
|
A. Okuta, K. Ohnishi, and S. Harayama Construction of Chimeric Catechol 2,3-Dioxygenase Exhibiting Improved Activity against the Suicide Inhibitor 4-Methylcatechol Appl. Envir. Microbiol., March 1, 2004; 70(3): 1804 - 1810. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Sondossi, D. Barriault, and M. Sylvestre Metabolism of 2,2'- and 3,3'-Dihydroxybiphenyl by the Biphenyl Catabolic Pathway of Comamonas testosteroni B-356 Appl. Envir. Microbiol., January 1, 2004; 70(1): 174 - 181. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. E. Parales, N. C. Bruce, A. Schmid, and L. P. Wackett Biodegradation, Biotransformation, and Biocatalysis (B3) Appl. Envir. Microbiol., October 1, 2002; 68(10): 4699 - 4709. [Full Text] [PDF]
|
|
|
|
|
|
Y. Ge, F. H. Vaillancourt, N. Y. R. Agar, and L. D. Eltis Reactivity of Toluate Dioxygenase with Substituted Benzoates and Dioxygen J. Bacteriol., August 1, 2002; 184(15): 4096 - 4103. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E.-S. Mohamed, A. Zaar, C. Ebenau-Jehle, and G. Fuchs Reinvestigation of a New Type of Aerobic Benzoate Metabolism in the Proteobacterium Azoarcus evansii J. Bacteriol., March 15, 2001; 183(6): 1899 - 1908. [Abstract] [Full Text]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |