DIFFERENTIAL EXPRESSION OF ENZYME-ACTIVITIES INITIATING ANOXIC METABOLISM OF VARIOUS AROMATIC-COMPOUNDS VIA BENZOYL-COA

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 (CO2)-C-14 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.