| 177 | 0 | 24 |
| 下载次数 | 被引频次 | 阅读次数 |
茶黄素因其生理活性在医药和功能性食品领域备受关注。本研究对茶黄素的酶促合成进行了工艺优化，并利用大孔树脂对其进行纯化。通过单因素和响应面分析确定茶黄素的最佳酶促合成条件：50 mL茶汤添加40 mL酶液，29℃反应26 min，茶黄素合成量达（173.63±1.69）μg/mL，儿茶素转化率92.18%，合成率14.02%，相当于每克绿茶粉生成15.6 mg茶黄素。粗茶黄素粉纯度为（30.23%±2.09）%。XAD-16N树脂动态吸附参数：上样流速5 mL/min、质量浓度5 mg/mL，解吸梯度为3 BV水、6 BV 30%乙醇和6 BV 60%乙醇，纯化后茶黄素纯度提升2.92倍。该研究为茶黄素的酶法合成与树脂纯化提供了技术支持。
Abstract：Theaflavins have garnered significant attention in medicine and functional foods due to their physiological activities. In this study,the enzymatic synthesis process of theaflavins was optimized,and the theaflavin product was purified using macroporous resin. Optimal enzymatic synthesis conditions,determined through single-factor experiments and response surface methodology,were as following:reacting 50 mL of tea soup with 40 mL of enzyme solution,at 29 ℃ for 26 min. Under the above conditions,the theaflavin yield reached(173.63±1.69) μg/mL,with a catechin conversion rate of 92.18% and a synthesis rate of 14.02%,corresponding to 15.6 mg per gram of green tea powder. The purity of crude theaflavin powder was 30.23%±2.09%. For purification,XAD-16N resin was used with a sample loading flow rate of 5 mL/min and a concentration of 5 mg/mL. Desorption was achieved with a gradient of 3 bed volumes(BV) of water,followed by 6 BV of 30% ethanol and 6 BV of 60% ethanol. After purification,the theaflavin purity increased by 2.92 times. This research provides technical support for the enzymatic synthesis and resin-based purification of theaflavins.
[1]Takemoto M,Takemoto H. Synthesis of theaflavins and their functions[J]. Molecules,2018,23(4):918.
[2]Liu Y F,Wang D Y,Li J,et al. Research progress on the functions and biosynthesis of theaflavins[J]. Food Chemistry,2024,450:139285.
[3]Wu D,Chen R H,Zhang W J,et al. Tea and its components reduce the production of uric acid by inhibiting xanthine oxidase[J]. Food&Nutrition Research,2022,66:8239.
[4]Noor Mohammadi T,Maung A T,Sato J,et al. Mechanism for antibacterial action of epigallocatechin gallate and theaflavin-3,3’-digallate on Clostridium perfringens.[J].Journal of Applied Microbiology,2019,126(2):633-640.
[5]Mhatre S, Srivastava T, Naik S, et al. Antiviral activity of green tea and black tea polyphenols in prophylaxis and treatment of COVID-19:A review[J]. Phytomedicine,2021,85:153286.
[6]Zhang Y H,Liu W L,Wang X Y,et al. Nanozyme-enabled treatment of cardio-and cerebrovascular diseases[J]. Small,2023,19(13):2204809.
[7]Chen Y,Luo L Y,Hu S S,et al. The chemistry,processing,and preclinical anti-hyperuricemia potential of tea:A comprehensive review[J]. Critical Reviews in Food Science and Nutrition,2023,63(24):7065-7090.
[8]刘仲华，张盛，刘昌伟，等.茶叶功能成分利用“十三五”进展及“十四五”发展方向[J].中国茶叶，2021, 43(10):1-9.
[9]Liu Q,Xun G H,Feng Y. The state-of-the-art strategies of protein engineering for enzyme stabilization[J]. Biotechnology Advances,2019,37(4):530-537.
[10]Subramanian N,Venkatesh P,Ganguli S,et al. Role of polyphenol oxidase and peroxidase in the generation of black tea theaflavins[J]. Journal of Agricultural and Food Chemistry,1999,47(7):2571-2578.
[11]谷记平，刘仲华，黄建安，等.酶性氧化合成茶黄素条件优化的研究[J].茶叶科学，2006,26(4):285-290.
[12]Cai H L,Zhong Z H,Chen Y R,et al. Genes cloning,sequencing and function identification of recombinant polyphenol oxidase isozymes for production of monomeric theaflavins from Camellia sinensis[J]. International Journal of Biological Macromolecules,2023,240:124353.
[13]Teng J,Liu Y,Zeng W,et al. In vitro enzymatic synthesis of a monomeric theaflavin using a polyphenol oxidase isozyme from tea(Camellia sinensis)leaf[J]. International Journal of Food Science&Technology,2022,57(9):5621-5631.
[14]Li D, Dong L, Li J Y, et al. Optimization of enzymatic synthesis of theaflavins from potato polyphenol oxidase[J]. Bioprocess and Biosystems Engineering, 2022, 45(6):1047-1055.
[15]Lei S C,Xie M H,Hu B,et al. Effective synthesis of theaflavin-3,3'-digallate with epigallocatechin-3-O-gallate and epicatechin gallate as substrates by using immobilized pear polyphenol oxidase[J]. International Journal of Biological Macromolecules,2017,94(Pt A):709-718.
[16]Zeng J, Du G, Shao X, et al. Recombinant polyphenol oxidases for production of theaflavins from tea polyphenols[J].International Journal of Biological Macromolecules, 2019,134:139-145.
[17]国家标准化管理委员会，国家质量监督检验检疫总局.茶叶中茶黄素的测定-高效液相色谱法：GB/T 30483-2013[S].北京：中国标准出版社，2014.
[18]张颖.酶促氧化制备茶黄素及其性质研究[D].天津：天津科技大学，2010.
[19]王坤波，刘仲华，赵淑娟，等.儿茶素组成和理化条件对茶黄素酶催化合成的影响[J].茶叶科学，2007, 27(3):192-200.
[20]Xu J,Wei Y,Huang Y,et al. Current understanding and future perspectives on the extraction,structures,and regulation of muscle function of tea pigments[J]. Critical Reviews in Food Science and Nutrition,2023,63(33):11522-11544.
[21]李大祥，宛晓春，杨昌军，等.茶儿茶素氧化机理[J].天然产物研究与开发，2006,18(1):171-181.
[22]王坤波.茶黄素的酶促合成、分离鉴定及功能研究[D].长沙：湖南农业大学，2007.
[23]Sun H J, Wang J, Tao X M, et al. Purification and characterization of polyphenol oxidase from rape flower[J]. Journal of Agricultural and Food Chemistry, 2012, 60(3):823-829.
[24]Robertson A. Effects of physical and chemical conditions on the in vitro oxidation of tea leaf catechins[J]. Phytochemistry,1983,22(4):889-896.
[25]徐洪梅.梨多酚氧化酶生物催化合成茶黄素及其在茶叶深加工中的应用研究[D].杭州：浙江工业大学，2013.
[26]袁斌.茶黄素的酶促氧化制备与纯化技术研究[D].长沙：湖南农业大学，2020.
[27]Nazemi R,Ashoori S,Moghadasi J. Investigating the performance of non-damaging hybrid gel to mitigate lost circulation in fractured formations:A comprehensive study using Box–Behnken design[J]. Geoenergy Science and Engineering,2023,226:211730.
[28]Li Z P,Lu D G,Gao X J. Optimization of mixture proportions by statistical experimental design using response surface method-A review[J]. Journal of Building Engineering,2021,36:102101.
基本信息:
DOI：10.19804/j.issn1006-2513.2025.11.008
中图分类号:TS209
引用信息:
[1]李佳,毛立科,何涛,等.基于绿茶加工副产物的茶黄素酶法合成工艺优化研究[J].中国食品添加剂,2025,36(11):59-68.DOI:10.19804/j.issn1006-2513.2025.11.008.
基金信息: