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J Vis Exp. 2018 Oct 2;(140). doi: 10.3791/58166.

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions.

Author information

1
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán.
2
CONACYT, Facultad de Ingeniería Química, Campus de Ciencias Exactas e Ingeniería, Universidad Autónoma de Yucatán.
3
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán; ths@cicy.mx.

Abstract

Caffeine (1,3,7-trimethylxanthine) is a purine alkaloid present in popular drinks such as coffee and tea. This secondary metabolite is regarded as a chemical defense because it has antimicrobial activity and is considered a natural insecticide. Caffeine can also produce negative allelopathic effects that prevent the growth of surrounding plants. In addition, people around the world consume caffeine for its analgesic and stimulatory effects. Due to interest in the technological applications of caffeine, research on the biosynthetic pathway of this compound has grown. These studies have primarily focused on understanding the biochemical and molecular mechanisms that regulate the biosynthesis of caffeine. In vitro tissue culture has become a useful system for studying this biosynthetic pathway. This article will describe a step-by-step protocol for the quantification of caffeine and for measuring the transcript levels of the gene (CCS1) encoding caffeine synthase (CS) in cell suspensions of C. arabica L. as well as its activity.

PMID:
30346406
PMCID:
PMC6235369
[Available on 2020-10-02]
DOI:
10.3791/58166
[Indexed for MEDLINE]

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