Eintrag weiter verarbeiten
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls
Gespeichert in:
| Zeitschriftentitel: | Journal of Bacteriology |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | Journal of Bacteriology, 183, 2001, 5, S. 1511-1516 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Seah, Stephen Y. K. Labbé, Geneviève Kaschabek, Stefan R. Reifenrath, Frank Reineke, Walter Eltis, Lindsay D. Seah, Stephen Y. K. Labbé, Geneviève Kaschabek, Stefan R. Reifenrath, Frank Reineke, Walter Eltis, Lindsay D. |
|---|---|
| author |
Seah, Stephen Y. K. Labbé, Geneviève Kaschabek, Stefan R. Reifenrath, Frank Reineke, Walter Eltis, Lindsay D. |
| spellingShingle |
Seah, Stephen Y. K. Labbé, Geneviève Kaschabek, Stefan R. Reifenrath, Frank Reineke, Walter Eltis, Lindsay D. Journal of Bacteriology Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls Molecular Biology Microbiology |
| author_sort |
seah, stephen y. k. |
| spelling |
Seah, Stephen Y. K. Labbé, Geneviève Kaschabek, Stefan R. Reifenrath, Frank Reineke, Walter Eltis, Lindsay D. 0021-9193 1098-5530 American Society for Microbiology Molecular Biology Microbiology http://dx.doi.org/10.1128/jb.183.5.1511-1516.2001 <jats:title>ABSTRACT</jats:title> <jats:p> 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (HOPDA) hydrolase (BphD) is a key determinant in the aerobic transformation of polychlorinated biphenyls (PCBs) by <jats:italic>Burkholderia</jats:italic> sp. strain LB400 (S. Y. K. Seah, G. Labbé, S. Nerdinger, M. Johnson, V. Snieckus, and L. D. Eltis, J. Biol. Chem. 275:15701–15708, 2000). To determine whether this is also true in divergent biphenyl degraders, the homologous hydrolase of <jats:italic>Rhodococcus globerulus</jats:italic> P6, BphD <jats:sub>P6</jats:sub> , was hyperexpressed, purified to apparent homogeneity, and studied by steady-state kinetics. BphD <jats:sub>P6</jats:sub> hydrolyzed HOPDA with a <jats:italic>k</jats:italic> <jats:sub>cat</jats:sub> / <jats:italic> K <jats:sub>m</jats:sub> </jats:italic> of 1.62 (± 0.03) × 10 <jats:sup>7</jats:sup> M <jats:sup>−1</jats:sup> s <jats:sup>−1</jats:sup> (100 mM phosphate [pH 7.5], 25°C), which is within 70% of that of BphD <jats:sub>LB400</jats:sub> . BphD <jats:sub>P6</jats:sub> was also similar to BphD <jats:sub>LB400</jats:sub> in that it catalyzed the hydrolysis of HOPDAs bearing chloro substituents on the phenyl moiety at least 25 times more specifically than those bearing chloro substituents on the dienoate moiety. However, the rhodococcal enzyme was significantly more specific for 9-Cl and 10-Cl HOPDAs, catalyzing the hydrolysis of 9-Cl, 10-Cl, and 9,10-diCl HOPDAs two- to threefold respectively, more specifically than HOPDA. Moreover, 4-Cl HOPDA competitively inhibited BphD <jats:sub>P6</jats:sub> more effectively than 3-Cl HOPDA, which is the inverse of what was observed in BphD <jats:sub>LB400</jats:sub> . These results demonstrate that BphD is a key determinant in the aerobic transformation of PCBs by divergent biphenyl degraders, but that there exists significant diversity in the specificity of these biphenyl hydrolases. </jats:p> Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls Journal of Bacteriology |
| doi_str_mv |
10.1128/jb.183.5.1511-1516.2001 |
| facet_avail |
Online Free |
| finc_class_facet |
Biologie |
| format |
ElectronicArticle |
| fullrecord |
blob:ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTEyOC9qYi4xODMuNS4xNTExLTE1MTYuMjAwMQ |
| id |
ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTEyOC9qYi4xODMuNS4xNTExLTE1MTYuMjAwMQ |
| institution |
DE-D275 DE-Bn3 DE-Brt1 DE-Zwi2 DE-D161 DE-Gla1 DE-Zi4 DE-15 DE-Pl11 DE-Rs1 DE-105 DE-14 DE-Ch1 DE-L229 |
| imprint |
American Society for Microbiology, 2001 |
| imprint_str_mv |
American Society for Microbiology, 2001 |
| issn |
0021-9193 1098-5530 |
| issn_str_mv |
0021-9193 1098-5530 |
| language |
English |
| mega_collection |
American Society for Microbiology (CrossRef) |
| match_str |
seah2001comparativespecificitiesoftwoevolutionarilydivergenthydrolasesinvolvedinmicrobialdegradationofpolychlorinatedbiphenyls |
| publishDateSort |
2001 |
| publisher |
American Society for Microbiology |
| recordtype |
ai |
| record_format |
ai |
| series |
Journal of Bacteriology |
| source_id |
49 |
| title |
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_unstemmed |
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_full |
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_fullStr |
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_full_unstemmed |
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_short |
Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_sort |
comparative specificities of two evolutionarily divergent hydrolases involved in microbial degradation of polychlorinated biphenyls |
| topic |
Molecular Biology Microbiology |
| url |
http://dx.doi.org/10.1128/jb.183.5.1511-1516.2001 |
| publishDate |
2001 |
| physical |
1511-1516 |
| description |
<jats:title>ABSTRACT</jats:title>
<jats:p>
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (HOPDA) hydrolase (BphD) is a key determinant in the aerobic transformation of polychlorinated biphenyls (PCBs) by
<jats:italic>Burkholderia</jats:italic>
sp. strain LB400 (S. Y. K. Seah, G. Labbé, S. Nerdinger, M. Johnson, V. Snieckus, and L. D. Eltis, J. Biol. Chem. 275:15701–15708, 2000). To determine whether this is also true in divergent biphenyl degraders, the homologous hydrolase of
<jats:italic>Rhodococcus globerulus</jats:italic>
P6, BphD
<jats:sub>P6</jats:sub>
, was hyperexpressed, purified to apparent homogeneity, and studied by steady-state kinetics. BphD
<jats:sub>P6</jats:sub>
hydrolyzed HOPDA with a
<jats:italic>k</jats:italic>
<jats:sub>cat</jats:sub>
/
<jats:italic>
K
<jats:sub>m</jats:sub>
</jats:italic>
of 1.62 (± 0.03) × 10
<jats:sup>7</jats:sup>
M
<jats:sup>−1</jats:sup>
s
<jats:sup>−1</jats:sup>
(100 mM phosphate [pH 7.5], 25°C), which is within 70% of that of BphD
<jats:sub>LB400</jats:sub>
. BphD
<jats:sub>P6</jats:sub>
was also similar to BphD
<jats:sub>LB400</jats:sub>
in that it catalyzed the hydrolysis of HOPDAs bearing chloro substituents on the phenyl moiety at least 25 times more specifically than those bearing chloro substituents on the dienoate moiety. However, the rhodococcal enzyme was significantly more specific for 9-Cl and 10-Cl HOPDAs, catalyzing the hydrolysis of 9-Cl, 10-Cl, and 9,10-diCl HOPDAs two- to threefold respectively, more specifically than HOPDA. Moreover, 4-Cl HOPDA competitively inhibited BphD
<jats:sub>P6</jats:sub>
more effectively than 3-Cl HOPDA, which is the inverse of what was observed in BphD
<jats:sub>LB400</jats:sub>
. These results demonstrate that BphD is a key determinant in the aerobic transformation of PCBs by divergent biphenyl degraders, but that there exists significant diversity in the specificity of these biphenyl hydrolases.
</jats:p> |
| container_issue |
5 |
| container_start_page |
1511 |
| container_title |
Journal of Bacteriology |
| container_volume |
183 |
| format_de105 |
Article, E-Article |
| format_de14 |
Article, E-Article |
| format_de15 |
Article, E-Article |
| format_de520 |
Article, E-Article |
| format_de540 |
Article, E-Article |
| format_dech1 |
Article, E-Article |
| format_ded117 |
Article, E-Article |
| format_degla1 |
E-Article |
| format_del152 |
Buch |
| format_del189 |
Article, E-Article |
| format_dezi4 |
Article |
| format_dezwi2 |
Article, E-Article |
| format_finc |
Article, E-Article |
| format_nrw |
Article, E-Article |
| _version_ |
1792341550500413440 |
| geogr_code |
not assigned |
| last_indexed |
2024-03-01T16:21:42.512Z |
| geogr_code_person |
not assigned |
| openURL |
url_ver=Z39.88-2004&ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fvufind.svn.sourceforge.net%3Agenerator&rft.title=Comparative+Specificities+of+Two+Evolutionarily+Divergent+Hydrolases+Involved+in+Microbial+Degradation+of+Polychlorinated+Biphenyls&rft.date=2001-03-01&genre=article&issn=1098-5530&volume=183&issue=5&spage=1511&epage=1516&pages=1511-1516&jtitle=Journal+of+Bacteriology&atitle=Comparative+Specificities+of+Two+Evolutionarily+Divergent+Hydrolases+Involved+in+Microbial+Degradation+of+Polychlorinated+Biphenyls&aulast=Eltis&aufirst=Lindsay+D.&rft_id=info%3Adoi%2F10.1128%2Fjb.183.5.1511-1516.2001&rft.language%5B0%5D=eng |
| SOLR | |
| _version_ | 1792341550500413440 |
| author | Seah, Stephen Y. K., Labbé, Geneviève, Kaschabek, Stefan R., Reifenrath, Frank, Reineke, Walter, Eltis, Lindsay D. |
| author_facet | Seah, Stephen Y. K., Labbé, Geneviève, Kaschabek, Stefan R., Reifenrath, Frank, Reineke, Walter, Eltis, Lindsay D., Seah, Stephen Y. K., Labbé, Geneviève, Kaschabek, Stefan R., Reifenrath, Frank, Reineke, Walter, Eltis, Lindsay D. |
| author_sort | seah, stephen y. k. |
| container_issue | 5 |
| container_start_page | 1511 |
| container_title | Journal of Bacteriology |
| container_volume | 183 |
| description | <jats:title>ABSTRACT</jats:title> <jats:p> 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (HOPDA) hydrolase (BphD) is a key determinant in the aerobic transformation of polychlorinated biphenyls (PCBs) by <jats:italic>Burkholderia</jats:italic> sp. strain LB400 (S. Y. K. Seah, G. Labbé, S. Nerdinger, M. Johnson, V. Snieckus, and L. D. Eltis, J. Biol. Chem. 275:15701–15708, 2000). To determine whether this is also true in divergent biphenyl degraders, the homologous hydrolase of <jats:italic>Rhodococcus globerulus</jats:italic> P6, BphD <jats:sub>P6</jats:sub> , was hyperexpressed, purified to apparent homogeneity, and studied by steady-state kinetics. BphD <jats:sub>P6</jats:sub> hydrolyzed HOPDA with a <jats:italic>k</jats:italic> <jats:sub>cat</jats:sub> / <jats:italic> K <jats:sub>m</jats:sub> </jats:italic> of 1.62 (± 0.03) × 10 <jats:sup>7</jats:sup> M <jats:sup>−1</jats:sup> s <jats:sup>−1</jats:sup> (100 mM phosphate [pH 7.5], 25°C), which is within 70% of that of BphD <jats:sub>LB400</jats:sub> . BphD <jats:sub>P6</jats:sub> was also similar to BphD <jats:sub>LB400</jats:sub> in that it catalyzed the hydrolysis of HOPDAs bearing chloro substituents on the phenyl moiety at least 25 times more specifically than those bearing chloro substituents on the dienoate moiety. However, the rhodococcal enzyme was significantly more specific for 9-Cl and 10-Cl HOPDAs, catalyzing the hydrolysis of 9-Cl, 10-Cl, and 9,10-diCl HOPDAs two- to threefold respectively, more specifically than HOPDA. Moreover, 4-Cl HOPDA competitively inhibited BphD <jats:sub>P6</jats:sub> more effectively than 3-Cl HOPDA, which is the inverse of what was observed in BphD <jats:sub>LB400</jats:sub> . These results demonstrate that BphD is a key determinant in the aerobic transformation of PCBs by divergent biphenyl degraders, but that there exists significant diversity in the specificity of these biphenyl hydrolases. </jats:p> |
| doi_str_mv | 10.1128/jb.183.5.1511-1516.2001 |
| facet_avail | Online, Free |
| finc_class_facet | Biologie |
| format | ElectronicArticle |
| format_de105 | Article, E-Article |
| format_de14 | Article, E-Article |
| format_de15 | Article, E-Article |
| format_de520 | Article, E-Article |
| format_de540 | Article, E-Article |
| format_dech1 | Article, E-Article |
| format_ded117 | Article, E-Article |
| format_degla1 | E-Article |
| format_del152 | Buch |
| format_del189 | Article, E-Article |
| format_dezi4 | Article |
| format_dezwi2 | Article, E-Article |
| format_finc | Article, E-Article |
| format_nrw | Article, E-Article |
| geogr_code | not assigned |
| geogr_code_person | not assigned |
| id | ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTEyOC9qYi4xODMuNS4xNTExLTE1MTYuMjAwMQ |
| imprint | American Society for Microbiology, 2001 |
| imprint_str_mv | American Society for Microbiology, 2001 |
| institution | DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229 |
| issn | 0021-9193, 1098-5530 |
| issn_str_mv | 0021-9193, 1098-5530 |
| language | English |
| last_indexed | 2024-03-01T16:21:42.512Z |
| match_str | seah2001comparativespecificitiesoftwoevolutionarilydivergenthydrolasesinvolvedinmicrobialdegradationofpolychlorinatedbiphenyls |
| mega_collection | American Society for Microbiology (CrossRef) |
| physical | 1511-1516 |
| publishDate | 2001 |
| publishDateSort | 2001 |
| publisher | American Society for Microbiology |
| record_format | ai |
| recordtype | ai |
| series | Journal of Bacteriology |
| source_id | 49 |
| spelling | Seah, Stephen Y. K. Labbé, Geneviève Kaschabek, Stefan R. Reifenrath, Frank Reineke, Walter Eltis, Lindsay D. 0021-9193 1098-5530 American Society for Microbiology Molecular Biology Microbiology http://dx.doi.org/10.1128/jb.183.5.1511-1516.2001 <jats:title>ABSTRACT</jats:title> <jats:p> 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (HOPDA) hydrolase (BphD) is a key determinant in the aerobic transformation of polychlorinated biphenyls (PCBs) by <jats:italic>Burkholderia</jats:italic> sp. strain LB400 (S. Y. K. Seah, G. Labbé, S. Nerdinger, M. Johnson, V. Snieckus, and L. D. Eltis, J. Biol. Chem. 275:15701–15708, 2000). To determine whether this is also true in divergent biphenyl degraders, the homologous hydrolase of <jats:italic>Rhodococcus globerulus</jats:italic> P6, BphD <jats:sub>P6</jats:sub> , was hyperexpressed, purified to apparent homogeneity, and studied by steady-state kinetics. BphD <jats:sub>P6</jats:sub> hydrolyzed HOPDA with a <jats:italic>k</jats:italic> <jats:sub>cat</jats:sub> / <jats:italic> K <jats:sub>m</jats:sub> </jats:italic> of 1.62 (± 0.03) × 10 <jats:sup>7</jats:sup> M <jats:sup>−1</jats:sup> s <jats:sup>−1</jats:sup> (100 mM phosphate [pH 7.5], 25°C), which is within 70% of that of BphD <jats:sub>LB400</jats:sub> . BphD <jats:sub>P6</jats:sub> was also similar to BphD <jats:sub>LB400</jats:sub> in that it catalyzed the hydrolysis of HOPDAs bearing chloro substituents on the phenyl moiety at least 25 times more specifically than those bearing chloro substituents on the dienoate moiety. However, the rhodococcal enzyme was significantly more specific for 9-Cl and 10-Cl HOPDAs, catalyzing the hydrolysis of 9-Cl, 10-Cl, and 9,10-diCl HOPDAs two- to threefold respectively, more specifically than HOPDA. Moreover, 4-Cl HOPDA competitively inhibited BphD <jats:sub>P6</jats:sub> more effectively than 3-Cl HOPDA, which is the inverse of what was observed in BphD <jats:sub>LB400</jats:sub> . These results demonstrate that BphD is a key determinant in the aerobic transformation of PCBs by divergent biphenyl degraders, but that there exists significant diversity in the specificity of these biphenyl hydrolases. </jats:p> Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls Journal of Bacteriology |
| spellingShingle | Seah, Stephen Y. K., Labbé, Geneviève, Kaschabek, Stefan R., Reifenrath, Frank, Reineke, Walter, Eltis, Lindsay D., Journal of Bacteriology, Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls, Molecular Biology, Microbiology |
| title | Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_full | Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_fullStr | Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_full_unstemmed | Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_short | Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| title_sort | comparative specificities of two evolutionarily divergent hydrolases involved in microbial degradation of polychlorinated biphenyls |
| title_unstemmed | Comparative Specificities of Two Evolutionarily Divergent Hydrolases Involved in Microbial Degradation of Polychlorinated Biphenyls |
| topic | Molecular Biology, Microbiology |
| url | http://dx.doi.org/10.1128/jb.183.5.1511-1516.2001 |