SPECTROPHOTOMETRIC ASSAY FOR TRANS-CINNAMIC ACID 4-HYDROXYLASE ACTIVITY
SPECTROPHOTOMETRIC ASSAY FOR TRANS-CINNAMIC ACID 4-HYDROXYLASE ACTIVITY
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DOI：
10.1016/0003-2697(75)90651-x
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发表时间：
1975-01-01
影响因子：
2.9
通讯作者：
RUBERY, PH
中科院分区：
文献类型：
--
作者：
LAMB, CJ;RUBERY, PH
反式肉桂酸4-羟化酶（trans-Cinnamic acid, NADPH: oxygen oxidoreductase [4-hydroxylating]）可以通过分光光度法测定反应混合物酸化后，调整pH为11，在340 nm处产生4-羟基反式肉桂酸，从而快速准确地测定。在340 nm处测定粗提物和其他高本征吸收的制剂时，可以在分光光度测定前用乙醚将酸化后的测定混合物中的4-羟基反式肉桂酸提取到碱中进行修饰。反式肉桂酸2-羟化酶可在同一提取物中常规检测和测定。
trans-Cinnamic acid 4-hydroxylase (trans-cinnamic acid, NADPH: oxygen oxidoreductase [4-hydroxylating]) can be rapidly and precisely assayed by the spectrophotometric measurement of the production of 4-hydroxy-trans-cinnamic acid at 340 nm after acidification of the reaction mixture and subsequent readjustment to pH 11. For the assay of crude extracts and other preparations with high intrinsic absorption at 340 nm, the assay can be modified by extraction of the 4-hydroxy-trans-cinnamic acid from the acidified assay mixture through diethyl ether into alkali before spectrophotometric estimation. trans-Cinnamic acid 2-hydroxylase can be routinely detected and assayed in the same extract.