Production of a chimeric enzyme tool associating the Trichoderma reesei swollenin with the Aspergillus niger feruloyl esterase A for release of ferulic acid.
The main goals of this work were to produce the fusion protein of the Trichoderma reesei swollenin I (SWOI) and Aspergillus niger feruloyl esterase A (FAEA) and to study the effect of the physical association of the fusion partners on the efficiency of the enzyme. The fusion protein was produced up to 25 mg l(-1) in the T. reesei strains Rut-C30 and CL847. In parallel, FAEA alone was produced for use as a control protein in application tests. Recombinant FAEA and SWOI-FAEA were purified to homogeneity and characterized. The biochemical and kinetic characteristics of the two recombinant proteins were found to be similar to those of native FAEA, except for the temperature stability and specific activity of the SWOI-FAEA. Finally, the SWOI-FAEA protein was tested for release of ferulic acid from wheat bran. A period of 24 h of enzymatic hydrolysis with the SWOI-FAEA improved the efficiency of ferulic acid release by 50% compared with the results obtained using the free FAEA and SWOI. Ferulic acid is used as an antioxidant and flavor precursor in the food and pharmaceutical industries. This is the first report of a potential application of the SWOI protein fused with an enzyme of industrial interest.
Levasseur
Anthony
A
UMR 1163 INRA/Universités de Provence et de la Méditerranée de Biotechnologie des Champignons Filamenteux, IFR-IBAIM, 163 avenue de Luminy, Case Postale 925, 13288 Marseille Cedex 09, France.
Saloheimo
Markku
M
Navarro
David
D
Andberg
Martina
M
Monot
Frédéric
F
Nakari-Setälä
Tiina
T
Asther
Marcel
M
Record
Eric
E
D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D003373:Coumaric Acids / Q000378:metabolism*; D004043:Dietary Fiber / Q000378:metabolism; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
D003373:Coumaric Acids; D004043:Dietary Fiber; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase