葡萄糖苷酶制备冷水溶萝卜花青素修饰物的工艺优化和表征  

Optimized Preparation and Characterization of Cold Water-Soluble Radish Anthocyanin Modified by Glycosidase

作　　者：江文[1] 周桢[1] 李晓辉[1] 周小华[1] 周志明[1] 刘兵 JIANG Wen;ZHOU Zhen;LI Xiaohui;ZHOU Xiaohua;ZHOU Zhiming;LIU Bing(School of Chemistry and Chemical Engineering,Chongqing University,Chongqing 401331,China;Guangzhou Liushun Biological Co.Ltd.,Guangzhou 510032,China)

机构地区：[1]重庆大学化学化工学院,重庆401331 [2]广州六顺生物科技有限公司,广东广州510032

出　　处：《食品科学》2018年第18期235-241,共7页Food Science

基　　金：国家自然科学基金青年科学基金项目(21206175;315014682);天津市科委工业生物技术项目(14ZCZDSY00066);中央高校基本科研业务费专项(106112017CDJXFLX0014)

摘　　要：用膜分离和XAD-4大孔吸附树脂分离得到纯化萝卜花青素,经红外光谱和质谱分析表明其主要成分为萝卜花青素双糖苷,在4℃冷水和65%乙醇溶液中的溶解率仅为46.43%和63.07%。以纯化萝卜花青素为起始原料,进行葡萄糖苷酶修饰,探讨p H值、葡萄糖苷酶/萝卜花青素质量比、反应温度对制备萝卜花青素酶修饰物的影响,确定了糖苷酶修饰萝卜花青素的工艺条件,并用正交试验法对其工艺条件进行优化。红外光谱和质谱分析表明,葡萄糖苷酶修饰萝卜花青素的水解反应具有高度专一性,萝卜花青素双糖苷中的糖苷键被成功水解,生成的萝卜花青素酶修饰物为萝卜花青素单糖苷。通过糖苷酶的修饰,萝卜花青素的冷水可溶性得到了明显地改善,修饰后的萝卜花青素在4℃冷水和65%乙醇溶液中的溶解率分别提高115.38%和58.55%。采用溴邻苯三酚红氧化法的抗氧化能力分析表明,通过糖苷酶的修饰,萝卜花青素抗氧化能力得到明显提高,萝卜花青素酶修饰物对羟自由基的清除能力明显高于萝卜花青素,但是略低于原花青素B2。而且萝卜花青素酶修饰物可抑制人体肝癌细胞MHCC97L的生长,用浓度为5~40μmol/L的萝卜花青素酶修饰物处理人体肝癌细胞MHCC97L 24~72 h,均可显著抑制该细胞的生长,抑制率随其剂量增加而增大。Radish anthocyanins(RA)were purified by ultrafiltration and XAD-4 macroporous adsorption resin.PA was identified to consist mainly of anthocyanin diglucoside by Fourier transform infrared spectroscopy(FT-IR)and mass spectrometry(MS),and its solubility in 4℃cold water and 65%ethanol were 46.43%and 63.07%,respectively.Then RA was modified with glucosaccharase and the effect of pH,enzyme-to-substrate ratio and reaction temperature on the modification efficiency was optimized by orthogonal array design method.FT-IR and MS analyses indicated that the glycosidic bond of RA was specifically hydrolyzed by glucosaccharase into anthocyanin monoglycosides.The solubility of the modified anthocyanins in cold water and 65%ethanol was improved by 115.38%and 58.55%,respectively compared to the native counterpart.Furthermore,the hydroxyl radical scavenging activity of the modified anthocyanins,as determined using bromopyrogallol red oxidation assay,was significantly higher than that of native anthocyanins,but was slightly lower than that of procyanidine B2.Cytotoxicity evaluation suggested that the modified anthocyanins at concentrations from 5 to 40μmol/L significantly inhibited the cancer cell growth of MHCC97L cells for 24 to 72 h in a concentration-dependent manner.

关 键 词：萝卜花青素 纯化 酶修饰 抗氧化能力 细胞生长抑制 

分 类 号：TS201.2[轻工技术与工程—食品科学]