Spectrophotometric assay and electrophoretic detection of trans-feruloyl esterase activity.
Wheat bran cell walls were subjected to mild acid hydrolysis and the major phenolic product was purified and identified as 5-O-(trans-feruloyl)-arabinofuranose. Sensitive continuous and stopped, microtiter plate-based spectrophotometric assays for trans-feruloyl esterase activity were developed using this compound as substrate. Procedures were also developed for the detection of trans-feruloyl esterase activities on gels following electrophoresis using this compound. These procedures are applicable to other natural feruloyl esters derived from plant cell walls by enzymatic hydrolysis. The extracellular trans-feruloyl esterases of Aspergillus niger 814 grown on 1% wheat bran were fractionated by anion-exchange chromatography and isoelectric focusing. These studies indicate that there are multiple forms of trans-feruloyl esterase but that most activity is associated with a major isozyme with a pI of 3.2.
McCallum
J A
JA
Botany Department, University of British Columbia, Vancouver, Canada.
Taylor
I E
IE
Towers
G H
GH
D001089:Arabinose / Q000031:analogs & derivatives / Q000302:isolation & purification; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002473:Cell Wall / Q000201:enzymology; D003373:Coumaric Acids / Q000302:isolation & purification; D004586:Electrophoresis / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D014908:Triticum / Q000201:enzymology / Q000648:ultrastructure
D003373:Coumaric Acids; D007527:Isoenzymes; C072516:5-O-feruloylarabinose; D001089:Arabinose; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase