食品科学 ›› 2018, Vol. 39 ›› Issue (14): 296-301.doi: 10.7506/spkx1002-6630-201814044

• 工艺技术 • 上一篇    下一篇

茶黄素双没食子酸酯的酶法合成条件优化与纯化

吴光亮,刘新涛,胡冰,孙怡,曾晓雄*   

  1. (南京农业大学食品科技学院,江苏?南京 210095)
  • 出版日期:2018-07-25 发布日期:2018-07-16
  • 基金资助:
    江苏省重点研发计划(现代农业)重点及面上项目(BE2017374);江苏高校优势学科建设工程资助项目(PAPD)

Optimization of Enzymatic Synthesis and Purification of Theaflavin-3,3’-O-digallate

WU Guangliang, LIU Xintao, HU Bing, SUN Yi, ZENG Xiaoxiong*   

  1. (College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China)
  • Online:2018-07-25 Published:2018-07-16

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%。

关键词: 茶黄素双没食子酸, 多酚氧化酶, 酶法合成, 纯化

Abstract: 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%.

Key words: theaflavin-3,3’-O-gallate (TFDG), polyphenol oxidase (PPO), enzymatic synthesis, purification

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