Purification and characterization of a chlorogenic acid hydrolase from Aspergillus niger catalysing the hydrolysis of chlorogenic acid |
| |
Authors: | Asther Michèle Estrada Alvarado Maria Isabel Haon Mireille Navarro David Asther Marcel Lesage-Meessen Laurence Record Eric |
| |
Affiliation: | 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. |
| |
Abstract: | 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. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|