Walton N J, Skinner S E, Robins R J, Rhodes M J
AFRC Institute of Food Research, Norwich Laboratory, United Kingdom.
Anal Biochem. 1987 Jun;163(2):482-8. doi: 10.1016/0003-2697(87)90252-1.
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.
描述了一种用于测定 strictosidine 合酶的分光光度法。用乙酸乙酯提取 strictosidine，当使用高底物浓度时，有机提取物用稀氨水洗涤以除去共提取的 secologanin；溶剂蒸发后，残留物用 5M H2SO4 加热 45 分钟并测量 A348 值。根据同样处理的标准品的响应计算 strictosidine 的产量。可以测定出最低产量为 10 - 25 nmol 的 strictosidine。使用培养的金鸡纳树细胞提取物对该测定法进行了验证。
