马铃薯多酚氧化酶酶学性质及酶促合成茶黄素能力对比Enzymatic Properties of Potato Polyphenol Oxidase and Comparison of Enzymatic Synthesis Ability of Theaflavins
李东,张诗琪,陶迎梅,郝旺,李静雅,李宇豪
摘要(Abstract):
多酚氧化酶(polyphenol oxidase, PPO)在儿茶素转化生成茶黄素(theaflavins, TFs)的过程中起着关键作用。在酶促制备茶黄素中,不同PPO对儿茶素的催化作用导致TFs的合成量有所差异。文章分别从5个品种(黄心、黑美人、大牛角、红玫瑰、大白花)马铃薯中对PPO进行提取,用于研究不同PPO酶学性质及茶黄素的酶促制备能力。结果表明,5个品种马铃薯PPO最适反应温度为30~40℃,最适pH为6.0~7.2,最适反应底物浓度为50~60 mmol/L,在此范围内马铃薯PPO均表现出较高酶活力。在5个品种中黄心马铃薯PPO虽酶活力低于大牛角PPO,但其酶蛋白含量最高且催化合成TFs能力最强,TFs含量为129.71μg/mL,同时生成最强单体TFDG含量显著高于其他4个品种,可作为制备茶黄素最佳酶源。对比不同来源马铃薯PPO可知其酶学性质及催化合成TFs的合成量存在差异,为马铃薯PPO分离纯化提供了适宜环境和工业化生产茶黄素的酶促原料提供了参考。
关键词(KeyWords): 马铃薯多酚氧化酶;茶黄素;酶促反应;酶学性质
基金项目(Foundation): 四川苗子工程项目(2021084);; 四川省科技厅项目(2017NFP1068,2018NZYF0116)
作者(Author): 李东,张诗琪,陶迎梅,郝旺,李静雅,李宇豪
参考文献(References):
- [1]YANG Z Y,JIE G L,DONG F,et al.Radical-scavenging abilities and antioxidant properties of theaflavins and their gallate esters in H2O2 -mediated oxidative damage system in the HPF-1 cells[J].Toxicology in Vitro,2008,22(5):1250-1256.
- [2]CARLONI P,TIANO L,PADELLA L,et al.Antioxidant activity of white,green and black tea obtained from the same tea cultivar[J].Food Research International,2013,53(2):900-908.
- [3]ILARIA P,MAURO S.Antioxidants from black and green tea:from dietary modulation of oxidative stress to pharmacological mechanisms[J].British Journal of Pharmacology,2017,174(11):1195-1208.
- [4]左灵妮,刘康,马晓勤,等.茶黄素通过调控miR-190表达对LPS诱导的结肠上皮细胞炎症损伤的影响[J].中国免疫学杂志,2021,37(17):2087-2092.
- [5]王洪新,孙军涛,吕文平,等.茶黄素的制备、分析、分离及功能活性研究进展[J].食品与生物技术学报,2011,30(1):12-19.
- [6]刘伟,周洁,龚正礼.茶黄素的功能活性研究进展[J].食品科学,2013,34(11):386-391.
- [7]TENG J,GONG Z H,DENG Y L,et al.Purification,characterization and enzymatic synthesis of theaflavins of polyphenol oxidase isozymes from tea leaf (Camellia sinensis)[J].LWT-Food Science and Technology,2017,84:263-270.
- [8]龚志华,陈栋,朱盛尧,等.酶促合成茶黄素的茶鲜叶酶源筛选[J].茶叶科学,2013,33(1):34-39.
- [9]MATSUO Y,TADAKUMA F,SHII T,et al.Selective oxidation of pyrogallol-type catechins with unripe fruit homogenate of Citrus unshiu and structural revision of oolongtheanins[J].Tetrahedron,2015,71(17):2540-2548.
- [10]TAKASHI T,CHIE M,KYOKO I,et al.Synthesis of theaflavin from epicatechin and epigallocatechin by plant homogenates and role of epicatechin quinone in the synthesis and degradation of theaflavin[J].Journal of Agricultural and Food Chemistry,2002,50(7):2142-2148.
- [11]李洁媛,李东,童凯,等.马铃薯多酚氧化酶的分离纯化、酶学性质及酶促合成茶黄素性能研究[J].食品与发酵工业,2021,47(11):26-31.
- [12]WESTHUIZEN M,STEENKAMP L,STEENKAMP P,et al.Alternative pathway implicated as an influencing factor in the synthesis of theaflavin[J].Biocatalysis and Biotransformation,2015,33(5):298-309.
- [13]CHOW Y,LOUARME L,BONAZZI C,et al.Apple polyphenoloxidase inactivation during heating in the presence of ascorbic acid and chlorogenic acid[J].Food Chemistry,2011,129(3):761-767.
- [14]蒋长兴,焦云鹏,熊清平,等.山药氧化酶特性及茶黄素的合成[J].江苏农业科学,2013,41(8):277-280.
- [15]林彩霞,杨静蕊,王光瑛,等.多酚氧化酶催化合成茶黄素研究[J].植物生理学报,2017,53(8):1359-1364.
- [16]THYGESEN P W,DRY I B,ROBINSON S P.Polyphenol oxidase in potato.A multigene family that exhibits differential expression patterns[J].Plant Physiology,1995,109(2):525-531.
- [17]SHU J,MICHAEL H P.The effect of p-coumaric acid on browning inhibition in potato polyphenol oxidase-catalyzed reaction mixtures[J].Foods,2022,11(4):577.
- [18]SHU J,MICHAEL H P.Role of ascorbic acid in the extraction and quantification of potato polyphenol oxidase activity[J].Foods,2021,10(10):2486.
- [19]崔晓颖,彭新颜,贺红军,等.芋头多酚氧化酶的分离纯化与酶学特性[J].食品科学,2021,42(12):107-115.
- [20]袁斌,周诗雨,刘昌伟,等.多酚氧化酶的提取工艺与活性测定研究[J].茶叶通讯,2020,47(2):291-296.
- [21]李玉花,蓝兴国,聂玉哲.蛋白质分析实验技术指南[M].北京:高等教育出版社,2011.
- [22]黄莹捷,伍梦瑶,姚燕妮,等.不同反应条件对勐库大叶种多酚氧化酶合成茶黄素的影响[J].食品科学,2017,38(22):54-59.
- [23]李俊,卢扬,刘永翔,等.不同品种马铃薯多酚氧化酶酶学特性对比[J].食品科技,2020,45(6):218-223.
- [24]朱新鹏,孙敏,高天丽.不同条件下不同品种马铃薯PPO、POD活性[J].江苏农业科学,2015,43(8):97-99.
- [25]龙峻瑶,黄丽,夏宁,等.六堡茶发酵前后多酚氧化酶的分离及酶学性质[J].食品科学,2022,43(6):112-117.
- [26]WANG F H,ZHOU H L,CHENG F Y,et al.Comparison of the characterization and the temperature/pressure stability of soluble and membrane-bound polyphenol oxidase from 'Lijiang' snow peach[J].LWT-Food Science and Technology,2021,146(4):111401.
- [27]陈盛虎.龙井43号茶树多酚氧化酶同工酶的分离纯化与PPO Ⅲ-2、PPO Ⅴ-3性质研究[D].武汉:华中农业大学,2016.
- [28]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.
- [29]刘洋,陈勤操,刘德春,等.茶叶多酚氧化酶三相分离纯化及酶学性质研究[J].食品工业科技,2022,43(9):119-124.
- [30]徐斌.不同体系条件下聚酯型儿茶素的形成特性及其与茶黄素的竞争性形成[D].北京:中国农业科学院,2015.
- [31]伍梦瑶,黄莹捷,姚燕妮,等.勐库大叶种茶树多酚氧化酶粗酶的酶学性质[J].浙江大学学报(农业与生命科学版),2017,43(5):579-588.
- [32]罗玲,王洪新,胡倩倩,等.几种复合天然多酚氧化酶氧化茶多酚的比较研究[J].食品与生物技术学报,2015,34(4):385-389.
- [33]梁燕.微生物和水果源的多酚氧化酶固定化合成茶黄素研究[D].杭州:浙江大学,2008.
- [34]王坤波,刘仲华,赵淑娟,等.多酚氧化酶同工酶组成对茶黄素合成的影响[J].农业现代化研究,2007(5):618-621.
- [35]王馨雨,杨绿竹,王婷,等.植物多酚氧化酶的生理功能、分离纯化及酶促褐变控制的研究进展[J].食品科学,2020,41(9):222-237.
- [36]许雷.茶树多酚氧化酶的提取、分离纯化及其部分酶性质研究[D].武汉:华中农业大学,2014.
- [37]周仲华,张达,倪贺,等.生物信息学筛选催化茶黄素合成的多酚氧化酶[J].华南师范大学学报(自然科学版),2017,49(3):59-67.