Enzymes of anaerobic metabolism of phenolic compounds. 4-Hydroxybenzoate-CoA ligase from a denitrifying Pseudomonas species.

The initial step of anaerobic 4-hydroxybenzoate and 3-hydroxybenzoate degradation was studied in a denitrifying Pseudomonas sp. 4'-Hydroxybenzoate and 3-hydroxybenzoate are converted into their coenzyme A (CoA) thioesters by two different specific coenzyme A ligases. 4-Hydroxybenzoate-CoA ligase (AMP-forming) was purified 350-fold. The ligase is active as a monomer of molecular mass 48 kDa, as determined by gel filtration and SDS/PAGE. At a pH optimum of 8.5, the apparent Km values for 4-hydroxybenzoate, ATP, and coenzyme A are 37 microM, 77 microM, and 125 microM, respectively. The enzyme reacts specifically with 4-hydroxybenzoate (100%) and 4-aminobenzoate (30%). Other analogues of benzoate, notably 3- or 2-hydroxybenzoate, are inactive, and 2,4-dihydroxybenzoate and 2-hydroxy-4-methylbenzoate act as competitive inhibitors (Ki = 1 microM). Polyclonal antibodies were raised and used in immunoblot assays to study the regulation of the expression of 4-hydroxybenzoate-CoA ligase. The ligase is synthesized when cells are grown anaerobically with 4-hydroxybenzoate, phenol, or p-cresol; phenol and p-cresol are degraded via 4-hydroxybenzoate. The enzyme is not present in cells grown aerobically with 4-hydroxybenzoate or anaerobically with benzoate or 4-hydroxyphenylacetate.

[1]  G. Fuchs,et al.  Enzymes of anaerobic metabolism of phenolic compounds. 4-Hydroxybenzoyl-CoA reductase (dehydroxylating) from a denitrifying Pseudomonas species. , 1993, European journal of biochemistry.

[2]  G. Fuchs,et al.  Purification and characterization of phenylacetate-coenzyme A ligase from a denitrifying Pseudomonas sp., an enzyme involved in the anaerobic degradation of phenylacetate , 1993, Archives of Microbiology.

[3]  D. Elder,et al.  Evidence for two differentially regulated phenylpropenoyl-coenzyme A synthetase activities in Rhodopseudomonas palustris , 1992 .

[4]  F. Löffler,et al.  Purification and properties of 4-halobenzoate-coenzyme A ligase from Pseudomonas sp. CBS3. , 1992, Biological chemistry Hoppe-Seyler.

[5]  G. Fuchs,et al.  Novel aerobic 2-aminobenzoate metabolism. Purification and characterization of 2-aminobenzoate-CoA ligase, localisation of the gene on a 8-kbp plasmid, and cloning and sequencing of the gene from a denitrifying Pseudomonas sp. , 1992, European journal of biochemistry.

[6]  D. Dunaway-Mariano,et al.  Isolation and characterization of the three polypeptide components of 4-chlorobenzoate dehalogenase from Pseudomonas sp. strain CBS-3. , 1992, Biochemistry.

[7]  G. Fuchs,et al.  Enzymatic reduction of benzoyl-CoA to alicyclic compounds, a key reaction in anaerobic aromatic metabolism. , 1992, European journal of biochemistry.

[8]  C. Harwood,et al.  Regulation of benzoate-CoA ligase in Rhodopseudomonas palustris , 1991 .

[9]  G. Fuchs,et al.  Purification and characterization of benzoate-coenzyme A ligase and 2-aminobenzoate-coenzyme A ligases from a denitrifying Pseudomonas sp , 1991, Journal of bacteriology.

[10]  F. Löffler,et al.  Dehalogenation of 4-chlorobenzoate by 4-chlorobenzoate dehalogenase from pseudomonas sp. CBS3: an ATP/coenzyme A dependent reaction. , 1991, Biochemical and biophysical research communications.

[11]  J. Luengo,et al.  Purification and biochemical characterization of phenylacetyl-CoA ligase from Pseudomonas putida. A specific enzyme for the catabolism of phenylacetic acid. , 1990, The Journal of biological chemistry.

[12]  G. Fuchs,et al.  Reductive dehydroxylation of 4‐hydroxybenzoyl‐CoA to benzoyl‐CoA in a denitrifying, phenol‐degrading Pseudomonas species , 1989, FEBS letters.

[13]  J. Gibson,et al.  Involvement of coenzyme A thioesters in anaerobic metabolism of 4-hydroxybenzoate by Rhodopseudomonas palustris , 1989, Journal of bacteriology.

[14]  J. Gibson,et al.  Purification and properties of benzoate-coenzyme A ligase, a Rhodopseudomonas palustris enzyme involved in the anaerobic degradation of benzoate , 1988, Journal of bacteriology.

[15]  A. Larsson,et al.  Immunization of mice and rabbits by intrasplenic deposition of nanogram quantities of protein attached to Sepharose beads or nitrocellulose paper strips. , 1987, Journal of immunological methods.

[16]  D. Grbìc-Galìc.  Fermentative and oxidative transformation of ferulate by a facultatively anaerobic bacterium isolated from sewage sludge , 1985, Applied and environmental microbiology.

[17]  D. Metcalfe,et al.  An avidin--biotin microELISA for rapid measurement of total and allergen-specific human IgE. , 1983, Journal of immunological methods.

[18]  J. Morrissey,et al.  Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. , 1981, Analytical biochemistry.

[19]  M. Reinhard,et al.  Methanogenic Decomposition of Ferulic Acid, a Model Lignin Derivative , 1980, Applied and environmental microbiology.

[20]  M. Zenk,et al.  Procedure for the enzymatic synthesis and isolation of cinnamoyl-CoA thiolesters using a bacterial system. , 1980, Analytical biochemistry.

[21]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[22]  J. Mieyal,et al.  Benzoyl and hydroxybenzoyl esters of coenzyme A. Ultraviolet characterization and reaction mechanisms. , 1974, The Journal of biological chemistry.

[23]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[24]  P. Dutton,et al.  The photometabolism of benzoic acid by Rhodopseudomonas palustris: a new pathway of aromatic ring metabolism. , 1968, The Biochemical journal.