Purification and characterization of a chlorogenic acid hydrolase from Aspergillus niger catalysing the hydrolysis of chlorogenic acid.
Among 15 Aspergillus strains, Aspergillus niger BRFM 131 was selected for its high chlorogenic acid hydrolase activity. The enzyme was purified and characterized with respect to its physico-chemical and kinetic properties. Four chromatographic steps were necessary to purify the protein to homogeneity with a recovery of 2%. Km of the chlorogenic acid hydrolase was estimated to be 10 microM against chlorogenic acid as substrate. Under native conditions, the protein presented a molecular mass of 170 kDa, and SDS-PAGE analysis suggested the presence of two identical 80 kDa subunits. Isoelectric point was 6.0; pH optimum for activity was determined to be 6.0 and temperature optima to be 55 degrees C. The N-terminal sequence did not present any homology with other cinnamoyl ester hydrolases previously described suggesting the purification of a new protein. The chlorogenic acid hydrolase was used successfully for the production of caffeic acid, which possesses strong antioxidant properties, from natural substrates specially rich in chlorogenic acid like apple marc and coffee pulp.
Asther
Michèle
M
UMR 1163 INRA/Université de Provence de Biotechnologie des Champignons Filamenteux, IFR 86-IBAIM, Universités de Provence et de la Méditerranée, ESIL, 163 avenue de Luminy, Case Postale 925, 13288 Marseille 09, France.
Estrada Alvarado
Maria Isabel
MI
Haon
Mireille
M
Navarro
David
D
Asther
Marcel
M
Lesage-Meessen
Laurence
L
Record
Eric
E
D001234:Aspergillus niger / Q000145:classification* / Q000201:enzymology*; D002109:Caffeic Acids / Q000138:chemical synthesis*; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000302:isolation & purification*; D002384:Catalysis; D002726:Chlorogenic Acid / Q000737:chemistry*; D004789:Enzyme Activation; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D013045:Species Specificity; D013379:Substrate Specificity; D013696:Temperature
D002109:Caffeic Acids; D002726:Chlorogenic Acid; D002265:Carboxylic Ester Hydrolases; C025374:chlorogenic acid esterase; C040048:caffeic acid