Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Law, A.; Boulanger, M.J.
    Defining a structural and kinetic rationale for paralogous copies of phenylacetate-CoA ligases from the cystic fibrosis pathogen Burkholderia cenocepacia J2315 (2011), J. Biol. Chem., 286, 15577-15585.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.2.1.30 gene paaK1, paaK1 is a single copy gene, recombinant expression of His-tagged isozyme PaaK1 in Escherichia coli strain BL21Star(DE3), expression of selenomethionine-labeled PaaK1 in Escherichia coli strain 834(DE3) Burkholderia cenocepacia
6.2.1.30 gene paaK2, recombinant expression of His-tagged isozyme PaaK2 in Escherichia coli strain BL21Star(DE3) Burkholderia cenocepacia

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.2.1.30 adenylated phenylacetate intermediate complex of purified recombinant His-tagged PaaK2, sitting drop vapor diffusion method, 12 mg/ml protein in 20 mM HEPES, pH 8.3, 10 mM NaCl, 3 mM 2-mercaptoethanol, 2% glycerol, 3 mM MgCl2, and 3 mM ATP, is mixed with 17% w/v PEG 6000, 0.1 M HEPES, pH 7.5, 0.1 M KCl, and 2.5% glycerol, X-ray diffraction structure determination and analysis at 1.90 A resolution Burkholderia cenocepacia
6.2.1.30 purified recombinant His-tagged selenomethionine derivative of PaaK1 in complex with ATP and adenylated phenylacetate intermediate complex of purified recombinant His-tagged PaaK1, sitting drop vapor diffusion method, 12 mg/ml protein in 20 mM HEPES, pH 8.3, 10 mM NaCl, 3 mM 2-mercaptoethanol, 2% glycerol, 3 mM MgCl2, and 3 mM ATP, is mixed with 20-25% PEG 3350, 200 mM potassium thiocyanate, and 5% glycerol. Crystals of PaaK1 are grown from 10% w/v PEG 8000, 0.1 M HEPES, pH 7.5, and 8.0% v/v ethylene glycol, X-ray diffraction structure determination and analysis at 1.6 A and 1.92 A resolution, respectively Burkholderia cenocepacia

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.2.1.30 additional information
-
 additional information isozyme PaaK1 possesses a lower Km for phenylacetic acid and is better able to accommodate 3' and 4' substitutions on the phenyl ring than isozyme PaaK2 Burkholderia cenocepacia
6.2.1.30 0.062
-
 phenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 0.15
-
 phenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 0.17
-
 3-hydroxyphenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 0.23
-
 3-Chlorophenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 0.54
-
 4-hydroxyphenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 0.87
-
 3-hydroxyphenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 1.5
-
 3-Chlorophenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.2.1.30 Mg2+ required, the beta- and gamma-phosphates of ATP coordinate a single Mg2+ ion that is further coordinated by four water molecules in an overall octahedral geometry Burkholderia cenocepacia

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.2.1.30 ATP + phenylacetate + CoA Burkholderia cenocepacia
-
 AMP + diphosphate + phenylacetyl-CoA
-
 ?
6.2.1.30 ATP + phenylacetate + CoA Burkholderia cenocepacia DSM 16553
-
 AMP + diphosphate + phenylacetyl-CoA
-
 ?
6.2.1.30 additional information Burkholderia cenocepacia isozyme PaaK1 displays a lower Km for phenylacetic acid and broader substrate specificity than isozyme PaaK2 ?
-
 ?
6.2.1.30 additional information Burkholderia cenocepacia DSM 16553 isozyme PaaK1 displays a lower Km for phenylacetic acid and broader substrate specificity than isozyme PaaK2 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.2.1.30 Burkholderia cenocepacia B4E7B5 gene paaK1
-
6.2.1.30 Burkholderia cenocepacia B4EL89 gene paaK2
-
6.2.1.30 Burkholderia cenocepacia DSM 16553 B4E7B5 gene paaK1
-
6.2.1.30 Burkholderia cenocepacia DSM 16553 B4EL89 gene paaK2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.2.1.30 recombinant His-tagged isozyme PaaK1 from Escherichia coli strain BL21Star(DE3) and selenomethionine-labeled His-tagged PaaK1 from Escherichia coli strain 834(DE3) by nickel affinity chromatography, gel filtration, and anion exchange chromatography Burkholderia cenocepacia
6.2.1.30 recombinant His-tagged isozyme PaaK2 from Escherichia coli strain BL21Star(DE3) by nickel affinity chromatography, gel filtration, and anion exchange chromatography Burkholderia cenocepacia

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.2.1.30 ATP + 3-chlorophenylacetate + CoA
-
 Burkholderia cenocepacia AMP + diphosphate + 3-chlorophenylacetyl-CoA
-
 ?
6.2.1.30 ATP + 3-chlorophenylacetate + CoA
-
 Burkholderia cenocepacia DSM 16553 AMP + diphosphate + 3-chlorophenylacetyl-CoA
-
 ?
6.2.1.30 ATP + 3-hydroxyphenylacetate + CoA
-
 Burkholderia cenocepacia AMP + diphosphate + 3-hydroxyphenylacetyl-CoA
-
 ?
6.2.1.30 ATP + 3-hydroxyphenylacetate + CoA
-
 Burkholderia cenocepacia DSM 16553 AMP + diphosphate + 3-hydroxyphenylacetyl-CoA
-
 ?
6.2.1.30 ATP + 4-hydroxyphenylacetate + CoA
-
 Burkholderia cenocepacia AMP + diphosphate + 4-hydroxyphenylacetyl-CoA
-
 ?
6.2.1.30 ATP + 4-hydroxyphenylacetate + CoA
-
 Burkholderia cenocepacia DSM 16553 AMP + diphosphate + 4-hydroxyphenylacetyl-CoA
-
 ?
6.2.1.30 ATP + phenylacetate + CoA
-
 Burkholderia cenocepacia AMP + diphosphate + phenylacetyl-CoA
-
 ?
6.2.1.30 ATP + phenylacetate + CoA
-
 Burkholderia cenocepacia DSM 16553 AMP + diphosphate + phenylacetyl-CoA
-
 ?
6.2.1.30 additional information isozyme PaaK1 displays a lower Km for phenylacetic acid and broader substrate specificity than isozyme PaaK2 Burkholderia cenocepacia ?
-
 ?
6.2.1.30 additional information no activity 4-hydroxyphenylacetate, 3,4-dihydroxyphenylacetate and 2-chloro-2-hydroxy-phenylacetate Burkholderia cenocepacia ?
-
 ?
6.2.1.30 additional information no activity with 3,4-dihydroxyphenylacetate and 2-chloro-2-hydroxy-phenylacetate Burkholderia cenocepacia ?
-
 ?
6.2.1.30 additional information isozyme PaaK1 displays a lower Km for phenylacetic acid and broader substrate specificity than isozyme PaaK2 Burkholderia cenocepacia DSM 16553 ?
-
 ?
6.2.1.30 additional information no activity with 3,4-dihydroxyphenylacetate and 2-chloro-2-hydroxy-phenylacetate Burkholderia cenocepacia DSM 16553 ?
-
 ?
6.2.1.30 additional information no activity 4-hydroxyphenylacetate, 3,4-dihydroxyphenylacetate and 2-chloro-2-hydroxy-phenylacetate Burkholderia cenocepacia DSM 16553 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.2.1.30 PaaK1
-
 Burkholderia cenocepacia
6.2.1.30 PaaK2
-
 Burkholderia cenocepacia

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.2.1.30 1.17
-
 4-hydroxyphenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 1.6
-
 3-Chlorophenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 1.67
-
 3-Chlorophenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 1.83
-
 3-hydroxyphenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 2.67
-
 3-hydroxyphenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 4.17
-
 phenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia
6.2.1.30 5
-
 phenylacetate pH and temperature not specified in the publication Burkholderia cenocepacia

Cofactor

EC Number Cofactor Comment Organism Structure
6.2.1.30 ATP
-
 Burkholderia cenocepacia

General Information

EC Number General Information Comment Organism
6.2.1.30 evolution PaaK1 and PaaK2 form a unique subgroup within the adenylate-forming enzyme superfamily Burkholderia cenocepacia
6.2.1.30 additional information adenylated phenylacetate intermediate complexes of PaaK1 and PaaK2 occur in distinct conformations, a N-terminal microdomain may serve to recruit subsequent phenylacetate enzymes, whereas a bifunctional role is proposed for the P-loop in stabilizing the C-terminal domain in conformation 2, an extended aryl binding pocket in PaaK1 contrasts with PaaK2, overview Burkholderia cenocepacia
6.2.1.30 additional information isozyme PaaK1 shows dynamic enzyme-substrate interactions. Adenylated phenylacetate intermediate complexes of PaaK1 and PaaK2 occur in distinct conformations, a N-terminal microdomain may serve to recruit subsequent phenylacetate enzymes, whereas a bifunctional role is proposed for the P-loop in stabilizing the C-terminal domain in conformation 2, an extended aryl binding pocket in PaaK1 contrasts with PaaK2, overview Burkholderia cenocepacia
6.2.1.30 physiological function the phenylacetic acid degradation pathway is the sole aerobic route for phenylacetic acid metabolism in bacteria and facilitates degradation of environmental pollutants such as styrene and ethylbenzene. The PAA pathway also is implicated in promoting Burkholderia cenocepacia infections in cystic fibrosis patients Burkholderia cenocepacia
6.2.1.30 physiological function the phenylacetic acid degradation pathway is the sole aerobic route for phenylacetic acid metabolism in bacteria and facilitates degradation of environmental pollutants such as styrene and ethylbenzene. The phenylacetic acid pathway also is implicated in promoting Burkholderia cenocepacia infections in cystic fibrosis patients Burkholderia cenocepacia