Related Experiment Videos
A spectrophotometric assay for strictosidine synthase.
N J Walton1, S E Skinner, R J Robins
1AFRC Institute of Food Research, Norwich Laboratory, United Kingdom.
Analytical Biochemistry
|June 1, 1987
Summary
This study details a new spectrophotometric assay for strictosidine synthase. The method accurately quantifies strictosidine production in plant cell extracts, enabling sensitive biochemical analysis.
Area of Science:
- Biochemistry
- Plant Sciences
- Analytical Chemistry
Background:
- Strictosidine is a key precursor in the biosynthesis of indole alkaloids.
- Accurate quantification of strictosidine is essential for understanding plant secondary metabolism and for potential pharmaceutical applications.
Purpose of the Study:
- To develop and validate a simple, sensitive spectrophotometric assay for strictosidine synthase activity.
- To enable the determination of strictosidine production in plant cell cultures.
Main Methods:
- Extraction of strictosidine using ethyl acetate.
- Removal of interfering secologanin with dilute ammonia.
- Acid hydrolysis of strictosidine with sulfuric acid.
- Spectrophotometric measurement of absorbance at 348 nm (A348).
Main Results:
- The assay allows for the determination of strictosidine production with a minimum quantifiable amount of 10-25 nmol.
- The method was successfully demonstrated using extracts from cultured Cinchona ledgeriana cells.
- The assay provides a reliable measure of strictosidine synthase activity.
Conclusions:
- A robust spectrophotometric assay for strictosidine synthase has been established.
- This assay is suitable for quantifying strictosidine production in plant extracts, particularly from cell cultures.
- The developed method offers a valuable tool for phytochemical research and drug discovery efforts.