Synthesis of 4-nitrophenyl caffeate and its use in assays of caffeoyl esterases

Anal Biochem. 2014 Jan 15:445:49-53. doi: 10.1016/j.ab.2013.10.006. Epub 2013 Oct 14.

Abstract

We have prepared 4-nitrophenyl caffeate by a combination of standard procedures of organic synthesis and enzymatic deacetylation. Based on hydrolysis of 4-nitrophenyl caffeate, a convenient spectrophotometric assay was developed for specific monitoring of caffeoyl esterase. The method is fast and easy to perform, and it requires no expensive equipment. Its reliability was tested on eight enzyme preparations comprising various combinations of caffeoyl, feruloyl, and acetyl esterase as well as protease activities.

Keywords: 4-Nitrophenyl caffeate; Caffeoyl esterase; HPLC assay; Spectrophotometric assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocatalysis
  • Caffeic Acids / chemical synthesis
  • Caffeic Acids / chemistry*
  • Caffeic Acids / metabolism
  • Carboxylic Ester Hydrolases / metabolism*
  • Enzyme Assays
  • Nitrophenols / chemistry*
  • Spectrophotometry*
  • Substrate Specificity

Substances

  • Caffeic Acids
  • Nitrophenols
  • 4-nitrophenyl
  • Carboxylic Ester Hydrolases
  • feruloyl esterase
  • caffeic acid