A new tannase substrate for spectrophotometric assay.
A new tannase substrate, protocatechuic acid p-nitrophenyl ester, 5, was synthesized using modern synthetic methods. The synthesis was designed to be performed by non-specialized chemists. It only involves four steps, three of which are protection-deprotection, and uses standard methods of separation and purification, such as recrystallization and column chromatography over silica. Under tannase action, protocatechuic acid p-nitrophenyl ester, 5, releases p-nitrophenol, which is easily measured spectrophotometrically either at 350 nm for pH values<6 or at 400 nm for pH values of 6-7 (yellow). The pH-response and the catalytic parameters of a crude Penicillium sp. tannase preparation were determined using 5 as substrate, thus showing the usefulness of this substrate in determining tannase activity.
Iacazio
G
G
Laboratoire de Microbiologie, Case 452, CNRS UPRESA 6116 Institut Méditerranéen d'Ecologie et de Paléoécologie, Faculté des Sciences de St Jérôme, Marseille, France. Gilles.Iacazio@Micro bio.U-3mrs.fr
Périssol
C
C
Faure
B
B
D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D062385:Hydroxybenzoates / Q000138:chemical synthesis* / Q000378:metabolism*; D007700:Kinetics; D009596:Nitrophenols; D010407:Penicillium / Q000201:enzymology; D013053:Spectrophotometry; D013056:Spectrophotometry, Ultraviolet
D062385:Hydroxybenzoates; D009596:Nitrophenols; C419872:protocatechuic acid 4-nitrophenyl ester; D002265:Carboxylic Ester Hydrolases; C022711:tannase