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Updated: May 6, 2026

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
Published on: October 2, 2018
Synthesis of 4-nitrophenyl caffeate and its use in assays of caffeoyl esterases
Andrej Chyba1, Vladimír Mastihuba, Mária Mastihubová
1Institute of Chemistry, Center for Glycomics, Slovak Academy of Sciences, SK-845 38 Bratislava, Slovakia.
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.
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