Position: Home > Articles > Optimization of Enzymatic Synthesis and Purification of Theaflavin-3,3'-O-digallate
FOOD SCIENCE
2018
(14)
296-301
茶黄素双没食子酸酯的酶法合成条件优化与纯化
作 者：
吴光亮;刘新涛;胡冰;孙怡;曾晓雄
单 位：
南京农业大学食品科技学院
关键词：
茶黄素双没食子酸;多酚氧化酶;酶法合成;纯化
摘 要：
利用皇冠梨多酚氧化酶催化表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)和表儿茶素没食子酸酯(epicatechin gallate,ECG)合成茶黄素双没食子酸酯(theaflavin-3,3’-O-digallate,TFDG),首先单因素试验确定了酶促反应的底物EGCG/ECG物质的量比3∶1、pH 4.0、反应温度35℃,在此基础上采用Box-Behnken试验设计法进行3因素3水平响应面优化试验,确定了TFDG最优酶促反应条件为底物EGCG/ECG物质的量比3.1∶1、pH 4.1、反应温度37℃;在最优反应条件下进行TFDG的合成,并利用AB-8层析柱结合高速逆流色谱技术分离纯化反应液,结果表明TFDG的得率和纯度分别达到85.14%和97%。
译 名：
Optimization of Enzymatic Synthesis and Purification of Theaflavin-3,3'-O-digallate
作 者：
WU Guangliang;LIU Xintao;HU Bing;SUN Yi;ZENG Xiaoxiong;College of Food Science and Technology, Nanjing Agricultural University;
单 位：
WU Guangliang%LIU Xintao%HU Bing%SUN Yi%ZENG Xiaoxiong%College of Food Science and Technology, Nanjing Agricultural University
关键词：
theaflavin-3,3'-O-gallate(TFDG);;polyphenol oxidase(PPO);;enzymatic synthesis;;purification
摘 要：
Polyphenol oxidase from Huangguan pear was applied to catalyze the synthesis of theaflavin-3,3'-O-gallate(TFDG) from epigallocatechin gallate(EGCG) and epicatechin gallate(ECG). By using one-factor-at-a-time method, the levels of EGCG/ECG molar ratio, pH and reaction temperature were selected in the range of 3:1, 4.0 and 35 ℃, respectively. These reaction conditions were optimized by response surface methodology using a three-variable, three-level Box-Behnken design. The optimal synthesis conditions for TFDG were determined as follows: EGCG/ECG molar ratio 3.1:1, pH 4.1, and 37 ℃. TFDG was obtained from the reaction mixture by AB-8 column chromatography and high-speed counter current chromatography with a yield of 85.14% and a purity of 97%.
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