Abstract
The regulation of the expression of enzyme activities catalyzing initial reactions in the anoxic metabolism of various aromatic compounds was studied at the whole cell level in the denitrifying Pseudomonas strain K 172. The specific enzyme activities were determined after growth on six different aromatic substrates (phenol, 4-hydroxybenzoate, benzoate, p-cresol, phenylacetate, 4-hydroxyphenylacetate) all being proposed to be metabolized anaerobically via benzoyl-CoA. As a control cells were grown on acetate, or aerobically on benzoate. The expression of the following enzyme activities was determined.
“Phenol carboxylase”, as studied by the isotope exchange between 14CO2 and the carboxyl group of 4-hydroxybenzoate; 4-hydroxybenzoyl-CoA reductase (dehydroxylating); p-cresol methylhydroxylase; 4-hydroxybenzyl alcohol dehydrogenase; 4-hydroxybenzaldehyde dehydrogenase; coenzymeA ligases for the aromatic acids benzoate, 4-hydroxybenzoate, phenylacetate, and 4-hydroxyphenylacetate; phenylglyoxylate: acceptor oxidoreductase and 4-hydroxyphenylglyoxylate: acceptor oxidoreductase; aromatic alcohol and aldehyde dehydrogenases.
The formation of most active enzymes is strictly regulated; they were only induced when required, the basic activities being almost zero. The observed whole cell regulation pattern supports the postulate that the enzyme activities play a role in anoxic aromatic metabolism and that the compounds are degraded via the following intermediates: Phenol → 4-hydroxybenzoate → 4-hydroxybenzoyl-CoA → benzoyl-CoA; 4-hydroxybenzoate → 4-hydroxybenzoyl-CoA → benzoyl-CoA; benzoate → benzoyl-CoA; p-cresol → 4-hydroxybenzaldehyde → 4-hydroxybenzoate → 4-hydroxybenzoyl-CoA → benzoyl-CoA; phenylacetate → phenylacetyl-CoA → phenylglyoxylate → benzoyl-CoA plus CO2; 4-hydroxyphenylacetate → 4-hydroxyphenylacetyl-CoA → 4-hydroxyphenylglyoxylate → 4-hydroxybenzoyl-CoA plus CO2 → benzoyl-CoA.
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Berry DF, Madsen EL, Bollag J-M (1987) Microbial metabolism of homocyclic and heterocyclic aromatic compounds under anaerobic conditions. Microbiol Rev 51:43–59
Blaschkowski HP, Neuer G, Ludwig-Fest LM, Knappe J (1982) Routes of flavodoxin and ferredoxin reduction in Escherichia coli. CoA-acetylating pyruvate: flavodoxin and NADPH: flavodoxin oxidoreductases participating in the activation of pyruvate formate-lyase. Eur J Biochem 123:563–569
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
Brune A, Schink B (1989) Pyrogallol-to-phloroglucinol conversion and other hydroxyl-transfer reactions catalyzed by cell extracts of Pelobacter acidigallici. J Bacteriol 172:1070–1076
Buchanan BB (1979) Ferredoxin-linked carbon dioxide fixation in photosynthetic bacteria. In: Gibbs M, Latzko E (eds) Photosynthesis. II. Photosynthetic carbon metabolism and related processes. Springer, Berlin Heidelberg New York, pp 416–424
Buder R, Fuchs G (1989) 2-Aminobenzoyl CoA monooxygenase/ reductase, a novel type of flavoenzyme. Purification and some properties of the enzyme. Eur J Biochem 185:629–635
Buder R, Ziegler K, Fuchs G, Langkau B, Ghisla S (1989) 2-Aminobenzoyl-CoA monooxygenase/reductase, a novel type of flavoenzyme. Studies on the stoichiometry and the course of the reaction. Eur J Biochem 185:637–643
Chalmers RM, Fewson CA (1989) Purification and characterization of benzaldehyde dehydrogenase I from Acinetobacter calcoaceticus. Biochem J 263:913–919
Evans WC, Fuchs G (1988) Anaerobic degradation of aromatic compounds. Annu Rev Microbiol 42:289–317
Gessler JF, Harwood CS, Gibson J (1988) Purification and properties of benzoate-coenzyme A ligase, a Rhodopseudomonas palustris enzyme involved in the anaerobic degradation of benzoate. J Bacteriol 170:1709–1714
Glöckler R, Tschech A, Fuchs G (1989) Reductive dehydroxylation of 4-hydroxybenzoyl-CoA to benzoyl-CoA in denitrifying, phenol-degrading Pseudomonas species. FEBS Lett 251:237–240
Goa J (1953) A microbiuret method for protein determination: determination of total protein in cerebrospinal fluid. Scand J Clin Lab Invest 5:128
Haddock JD, Ferry JG (1989) Purification and properties of phloroglucinol reductase from Eubacterium oxidoreducens G-41. J Biol Chem 264:4423–4427
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
Kerscher L, Oesterhelt D (1982) Pyruvate: ferredoxin oxidoreductase — new findings on an ancient enzyme. TIBS 371–374
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
Knoll G, Winter J (1989) Degradation of phenol via carboxylation to benzoate by a defined, obligate syntrophic consortium of anaerobic bacteria. Appl Microbiol Biotechnol 30:318–324
Koenig K, Andressen JR (1989) Molybdenum involvement in aerobic degradation of 2-furoic acid by Pseudomonas putida Fu1. Appl Environ Microbiol 55:1829–1834
Krumholz LR, Crawford RL, Hemling ME, Bryant MP (1987) Metabolism of gallate and phloroglucinol in Eubacterium oxidoreducens via 3-hydroxy-5-oxohexanoate. J Bacteriol 169:1886–1890
Krumholz LR, Bryant MP (1988) Characterization of the pyrogalol-phloroglucinol isomerase of Eubacterium oxidoreducens. J Bacteriol 170:2472–2479
Langkau B, Ghisla S, Buder R, Fuchs G (1990) 2-Aminobenzoyl-CoA monooxygenase/reductase, a novel type of flavoenzyme. Identification of the reaction products. Eur J Biochem 191:365–371
Martinez-Blanco H, Reglero A, Luengo JM (1990) Carbon catabolite regulation of phenylacetyl-CoA ligase from Pseudomonas putida. Biochem Biophys Res Commun 167:891–897
Merkel SM, Eberhard AE, Gibson J, Harwood CS (1989) Coenzyme A thioesters are involved in the anaerobic metabolism of 4-hydroxybenzoate by Rhodopseudomonas palustris. J Bacteriol 171:1–7
Müller R, Thiele J, Klages U, Lingens F (1984) Incorporation of [18O]water into 4-hydroxybenzoic acid in the reaction of 4-chlorobenzoate dehalogenase from Pseudomonas spec. CBS 3. Biochem Biophys Res Commun 124:178–182
Nozawa T, Maruyama Y (1988) Anaerobic metabolism of phthalate and other aromatic compounds by a denitrifying bacterium. J Bacteriol 170:5778–5784
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
Rudolphi A, Tschech A, Fuchs G (1991) Anaerobic degradation of cresols by denitrifying bacteria. Arch Microbiol 155:238–248
Schink B (1988) Principles and limits of anaerobic degradation: Environmental and technological aspects. In: Zehnder AJB (ed) Biology of anaerobic microorganisms. Wiley and Sons, New York, pp 771–846
Sembiring T, Winter J (1989) Anaerobic degradation of phenylacetic acid by mixed and pure cultures. Appl Microbiol Biotechnol 31:84–88
Seyfried B (1989) Anaerober Abbau von Phenylacetat über alpha-Oxidation durch nitratreduzierende Bakterien. Ph.D. thesis, University of Ulm
Seyfried B, Tschech A, Fuchs G (1991) Anaerobic degradation of phenylacetate and 4-hydroxyphenylacetate by denitrifying bacteria. Arch Microbiol 155:249–255
Trudgill PW (1984) The microbial metabolism of furans. In: Gibson DT (ed) Microbial degradation of organic compounds. Marcel Dekker, New York, pp 295–308
Tschech A (1989) Der anaerobe Abbau von aromatischen Verbindungen. Forum Mikrobiol 12:251–264
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
Tschech A, Schink B (1985) Fermentative degradation of resorcinol and resorcylic acids. Arch Microbiol 143:52–59
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 aerobic 2-aminobenzoate metabolism in a denitrifying Pseudomonas strain. Arch Microbiol 151:171–176
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Dangel, W., Brackmann, R., Lack, A. et al. Differential expression of enzyme activities initiating anoxic metabolism of various aromatic compounds via benzoyl-CoA. Arch. Microbiol. 155, 256–262 (1991). https://doi.org/10.1007/BF00252209
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DOI: https://doi.org/10.1007/BF00252209