An inducible hydrolase from Aspergillus niger, acting on carbon-carbon bonds, for phlorrhizin and other C-acylated phenols.
1. An inducible enzyme catalysing the hydrolysis of phloretin to form phloroglucinol and phloretic acid has been extracted from the acetone-dried powders of the mycelial felts of an Aspergillus niger strain grown in the presence of phlorrhizin. The enzyme was partially purified by treatment with protamine sulphate, ammonium sulphate fractionation, negative adsorption on tricalcium phosphate gel, and DEAE-cellulose column chromatography. 2. The hydrolytic activity on phloretin appeared to be maximal at about pH9.6. However, the characteristics of the enzyme were studied at pH7.2, because of the lability of the product, phloroglucinol, under alkaline conditions. 3. The apparent K(m) value at pH7.2 was about 0.3-0.4mm for phloretin and 0.15mm for 3'-methylphloracetophenone. 4. Maximum activity of the enzyme was obtained without the addition of any cofactor or metal ion. The involvement of thiol groups in the reaction was demonstrated by the potent inhibitory action of both heavy-metal ions and p-chloromercuribenzoate. 5. The enzyme showed a rather broad substrate specificity, and some other C-acylated phenols related to phloretin were hydrolysed. It was found that 3'-methylphloracetophenone, phloracetophenone and 2',4,4'-trihydroxydihydrochalcone were attacked more efficiently than phloretin. We propose the systematic name C-acylphenol acylhydrolase for the enzyme. This enzyme belongs to EC group 3.7.1.
D000098:Acetophenones / Q000378:metabolism; D001230:Aspergillus / Q000201:enzymology*; D002614:Chelating Agents / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose; D003470:Culture Media; D004790:Enzyme Induction; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000032:analysis* / Q000037:antagonists & inhibitors; D010693:Phloretin / Q000378:metabolism; D010695:Phlorhizin / Q000378:metabolism*
D000098:Acetophenones; D002614:Chelating Agents; D002729:Chloromercuribenzoates; D003470:Culture Media; D010695:Phlorhizin; D006867:Hydrolases; D010693:Phloretin