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. 1991 Aug 1;196(2):360-6.
doi: 10.1016/0003-2697(91)90479-d.

Spectrophotometric assay and electrophoretic detection of trans-feruloyl esterase activity

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Spectrophotometric assay and electrophoretic detection of trans-feruloyl esterase activity

J A McCallum et al. Anal Biochem. .

Abstract

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.

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