Abstract
The initial reactions in anaerobic metablism of methylphenols (cresols) and dimethylphenols were studied with denitrifying bacteria. A newly isolated strain, possibly a Paracoccus sp., was able to grow on o-or p-cresol as sole organic substrate with a generation time of 11 h; o-or p-cresol was completely oxidized to CO2 with nitrate being reduced to N2. A denitrifying Pseudomonas-like strain oxidized m-or p-cresol as the sole organic growth substrate completely to CO2 with a generation time of 14 h. Demonstration of intermediates and/or in vitro measurement of enzyme activities suggest the following enzymatic steps:
(1) p-Cresol was metabolized by both strains via benzoyl-CoA as central intermediate as follows: p-cresol → 4-OH-benzaldehyde → 4-OH-benzoate → 4-OH-benzoly-CoA → benzoyl-CoA. Oxidation of the methyl group to 4-OH-benzaldehyde was catalyzed by p-cresol methylhydroxylase. After oxidation of the aldehyde to 4-OH-benzoate, 4-OH-benzoyl-CoA is formed by 4-OH-benzoyl-CoA synthetase; subsequent reductive dehydroxylation of 4-OH-benzoyl-CoA to benzoyl-CoA is catalyzed by 4-OH-benzoyl-CoA reductase (dehydroxylating).
(2) o-Cresol was metabolized in the Paracoccus-like strain via 3-CH3-benzoyl-CoA as central intermediate as follows: o-cresol → 4-OH-3-CH3-benzoate → 4-OH-3-CH3-benzoyl-CoA → 3-CH3-benzoyl-CoA. The following enzymes were demonstrated: (a) An enzyme catalyzing an isototope exchange reaction between 14CO2 and the carboxyl of 4-OH-3-CH3-benzoate; this activity is thought to be a partial reaction catalyzed by an o-cresol carboxylase. (b) 4-OH-3-CH3-benzoyl-CoA synthetase (AMP-forming) activating the carboxylation product 4-OH-3-CH3-benzoate to its coenzyme A thioester. (c) 4-OH-3-CH3-benzoyl-CoA reductase (dehydroxylating) catalyzing the reductive dehydroxylation of the 4-hydroxyl group with reduced benzyl viologen as electron donor to yield 3-CH3-benzoyl-CoA. This thioester may also be formed by action of a coenzyme A ligase when 3-CH3-benzoate is metabolized. 2,4-Dimethylphenol was metabolized via 4-OH-3-CH3-benzoate and further to 3-CH3-benzoyl-CoA.
(3) The initial reactions of anaerobic metabolism of m-cresol in the Pseudomonas-like strain were not resolved. No indication for the oxidation of the methyl group nor for the carboxylation of m-cresol was found. In contrast, 2,4-and 3,4-dimethylphenol were oxidized to 4-OH-3-CH3-and 4-OH-2-CH3-benzoate, respectively, probably initiated by p-cresol methylhydroxylase; however, these compounds were not metabolized further.
Similar content being viewed by others
References
Bak F, Widdel F (1986) Anaerobic degradation of phenol and phenol derivatives by Desulfobacteium phenolicum sp. nov. Arch Microbiol 146: 177–180
Bakker G (1977) Anaerobic degradation of aromatic compounds in the presence of nitrate. FEMS Microbiol Lett 1: 103–108
Bayly RC, Dagley S, Gibson DT (1966) The metabolism of cresols by species of Pseudomonas, Biochem J 101: 293–301
Bossert ID, Young LY (1986) Anaerobic oxidation of p-cresol by a denitrifying bacterium. Appl Environ Microbiol 52: 1117–1122
Bossert ID, Whited G, Gibson DT, Young LY (1989) Anaerobic oxidation of p-cresol mediated by a partially purified methylhydroxylase from a denitrifying bacterium. J Bacteriol 171: 2956–2962
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254
Braunegg G, Sonnleitner B, Lafferty RM (1978) A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Eur J Appl Microbiol Biotechnol 6: 29–37
Chapman PJ, Hopper DJ (1968) The bacterial metabolism of 2,4-xylenol Biochem J 110: 491–498
Dagley S, Patel MD (1957) Oxidation of p-cresol and related compounds by a Pseudomonas. Biochem J 66: 227–233
Dangel W, Brackmann R, Lack A, Mohamed M, Koch J, Oswald B, Seyfried B, Tschech A, Fuchs G (1991) Differential expression of enzyme activities initiating anoxic metabolism of various aromatic compounds via benzoyl-CoA. Arch Microbiol 155: 256–262
Deley J (1970) Reexamination of the association between melting point, buoyant density and the chemical base composition of desoxyribonucleic acid. J Bacteriol 101: 738–754
Evans WC (1977) Biochemistry of the bacterial catabolism of aromatic compounds in anaerobic environments. Nature (Lond) 270: 17–22
Glöckler R, Tschech A, Fuchs G (1989) Reductive dehydroxylation of 4-hydroxybenzoyl-CoA to benzoyl-CoA in a denitrifying, phenol-degrading Pseudomonas species. FEBS Microbiol Lett 251: 237–240
Gregerson T (1978) Rapid method for distinction of gram-negative from gram-positive bacteria. Eur J Appl Microbiol Biotechnol 5: 123–127
Hopper DJ (1976) The hydroxylation of p-cresol and its conversion to p-hydroxybenzaldehyde in Pseudomonas putida. Biochem Biophys Res Commun 69: 462–468
Hopper DJ (1978) Incorporation of [18O]water in the formation of p-hydroxybenzyl alcohol by the p-cresol methylhydroxylase from Pseudomonas putida. Biochem J 175: 345–347
Hopper DJ, Taylor G (1975) Pathways for the degradation of m-cresol and p-cresol by Pseudomonas putida. J Bacteriol 122: 1–6
Kluge C, Tschech A, Fuchs G (1990) Anaerobic metabolism of resorcylic acids (m-dihydroxybenzoic acids) and resorcinol (1,3-benzenediol) in a fermentative and in a denitrifying bacterium. Arch Microbiol 155: 68–74
Magee CM, Rodeheaver G, Edgerton MT, Edlich RF (1975) A more reliable gram-staining technic for diagnosis of surgical infections. Am J Surg 130: 341–346
Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3: 208–218
McIntire W, Hopper DJ, Singer TP (1985) p-Cresol methylhydroxylase. Assay and general properties. Biochem J 228: 325–335
Merkel SM, Eberhard AE, Gibson J, Harwood CS (1989) Involvement of coenzyme A thioesters in anaerobic metabolism of 4-hydroxybenzoate by Rhodopseudomonas palustris. J Bacteriol 121: 1–7
Nozawa SM, Maruyama Y (1988) Anaerobic metabolism of phthalate and other aromatic compounds by a denitrifying bacterium. J Bacteriol 170: 5778–5784
Pfennig N, Wagener S (1986) An improved method of preparing wet mounts for photographs of microorganisms. J Microbiol Methods 4: 303–306
Rider BF, Mellon MG (1946) Colorimetric determination of nitrites. Ind Eng Chem 18: 96–98
Roberts DJ, Fedorak PM, Hrudey SE (1987) Comparison of the fates of the methyl carbons of m-cresol and p-cresol in methanogenic consortia. Can J Microbiol 33: 335–338
Roberts J, Fedorak PM, Hrudey SE (1990) CO2 incorporation and 4-hydroxy-2-methylbenzoic acid formation during anaerobic metabolism of m-cresol by a methanogenic consortium. Appl Environ Microbiol 56: 472–478
Schnell S, Bak F, Pfennig N (1989) Anaerobic degradation of aniline and dihydroxybenzenes by newly isolated sulfate-reducing bacteria and description of Desulfobacterium anilini. Arch Microbiol 152: 556–563
Smolenski WJ, Suflita JM (1987) Biodegradation of cresol isomers in anoxic aquifers. Appl Environ Microbiol 53: 710–716
Suflita WJ, Liang L, Saxena A (1989) The anaerobic biodegradation of o-, m-and p-cresol by sulfate-reducing bacterial enrichment cultures obtained from a shallow anoxic aquifer. J Ind Microbiol 4: 255–266
Tschech A, Fuchs G (1987) Anaerobic degradation of phenol by pure cultures of newly isolated denitrifying pseudomonads. Arch Microbiol 148: 213–217
Tschech A, Fuchs G (1989) Anaerobic degradation of phenol via carboxylation to 4-hydroxybenzoate: in vitro study of isotope exchange between 14CO2 and 4-hydroxybenzoate. Arch Microbiol 152: 594–599
Webster LT, Mieyal JJ, Siddiqui UA (1974) Benzoyl and hydroxybenzoyl esters of coenzyme A. Ultraviolet characterization and reaction mechanisms. J Biol Chem. 249: 2641–2645
Widdel F (1980) Anaerober Abbau von Fettsäuren und Benzoesäure durch neu isolierte Arten sulfatreduzierender Bakterien. Dissertation University Göttingen
Ziegler K, Braun K, Böckler A, Fuchs G (1987) Studies on the anaerobic degradation of benzoic acid and 2-aminobenzoic acid by a denitrifying Pseudomonas strain Arch Microbiol 149: 62–69
Ziegler K, Buder R, Winter J, Fuchs G (1989) Activation of aromatic acids and anaerobic 2-aminobenzoate metabolism in a denitrifying Pseudomonas strain. Arch Mirobiol 151: 171–176
Author information
Authors and Affiliations
Additional information
The hydroxyl and methyl groups are abbreviated as OH-and CH3-, respectively
Rights and permissions
About this article
Cite this article
Rudolphi, A., Tschech, A. & Fuchs, G. Anaerobic degradation of cresols by denitrifying bacteria. Arch. Microbiol. 155, 238–248 (1991). https://doi.org/10.1007/BF00252207
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00252207