A spectrophotometric assay for strictosidine synthase.
A spectrophotometric assay for strictosidine synthase is described. Strictosidine is extracted with ethyl acetate and, where high substrate concentrations are used, the organic extract is washed with dilute ammonia to remove coextracted secologanin; after evaporation of the solvent, the residue is heated with 5 M H2SO4 for 45 min and the A348 value is measured. Strictosidine production is calculated from the response of similarly treated standards. A minimum production of 10-25 nmol of strictosidine may be determined. The assay is demonstrated using extracts of cultured Cinchona ledgeriana cells.
Walton
N J
NJ
AFRC Institute of Food Research, Norwich Laboratory, United Kingdom.
Skinner
S E
SE
Robins
R J
RJ
Rhodes
M J
MJ
D019759:Carbon-Nitrogen Lyases; D002929:Cinchona / Q000201:enzymology; D026121:Indole Alkaloids; D057888:Iridoid Glucosides; D039823:Iridoids; D010946:Plants, Medicinal; D011714:Pyrans; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D014166:Transferases / Q000032:analysis*; D014363:Tryptamines; D014748:Vinca Alkaloids
D026121:Indole Alkaloids; D057888:Iridoid Glucosides; D039823:Iridoids; D011714:Pyrans; D014363:Tryptamines; D014748:Vinca Alkaloids; C021086:secologanin; C021093:strictosidine; C030820:tryptamine; D014166:Transferases; D019759:Carbon-Nitrogen Lyases; C022684:strictosidine synthetase