摘要
为探究工程酿酒酵母(Saccharomyces cerevisiae)发酵生产的番茄红素提取纯化方法及其结构,通过单因素和正交试验优化番茄红素的提取纯化工艺,并采用核磁共振(NMR)技术对其结构进行鉴定。结果表明,番茄红素的最佳提取工艺条件为酸热破壁:盐酸浓度3.0 mol/L,盐酸用量3.0 mL/0.1 g菌体,酸化时间60 min,酸化温度45℃;NaOH脱脂:NaOH浓度0.10 mol/L、NaOH用量2.0 mL/0.1 g菌体、皂化时间30 min,皂化后水浴温度40℃;丙酮提取∶料液比1∶20(g∶mL),提取温度40℃,提取时间50 min。最佳纯化工艺为正己烷析晶:40℃下用正己烷将粗提物溶解过滤,-20℃析晶12 h。在此优化提取纯化工艺条件下,番茄红素提取率为76%,纯度为96.7%。NMR结果显示,番茄红素结构为全反式番茄红素,与植物源番茄红素结构一致。
In order to explore the extraction and purification method and structure of lycopene produced by engineered Saccharomyces cerevisiae,the extraction and purification technology of lycopene was optimized by single factor and orthogonal tests,and its structure was identified by nuclear magnetic resonance(NMR).The results showed that the optimum extraction conditions of lycopene were acid heat breaking wall:hydrochloric acid concentration 3.0 mol/L,hydrochloric acid dosage 3.0 ml/0.1 g yeast,acidification time 60 min,and temperature 45℃.NaOH degreasing:NaOH concentration 0.10 mol/L,NaOH dosage 2.0 ml/0.1 g yeast,saponification time 30 min,water bath temperature 40℃after saponification.Acetone extraction∶material-liquid ratio 1∶20(g∶ml),extraction temperature 40℃,and time 50 min.The optimal purification process was n-hexane crystallization:crude extract was dissolved and filtered with n-hexane at 40℃and crystallized at-20℃for 12 h.Under the optimum extraction and purification conditions,the extraction rate of lycopene was 76%and the purity of lycopene was 96.7%.NMR results showed that the structure of lycopene was all-trans lycopene,which was consistent with plant-derived lycopene.
作者
石彬
朱伶俐
邓小敏
SHI Bin;ZHU Lingli;DENG Xiaomin(College of Life and Health,Wuhan Vocational College of Software and Engineering(Wuhan Open University),Wuhan 430205,China;Key Laboratory of Biology and Genetic Resources of Rubber Tree of Ministry of Agriculture and Rural Affairs State,Key Laboratory Incubation Base for Cultivation&Physiology of Tropical Crops,Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China)
出处
《中国酿造》
CAS
北大核心
2024年第8期202-208,共7页
China Brewing
基金
海南省自然科学基金高层次人才项目(320RC734)
武汉市知识创新项目曙光计划(2022022020801020440)
武汉软件工程职业学院科研启动基金(KYQDJF2023011)。
关键词
工程酿酒酵母
番茄红素
提取纯化
工艺优化
结构鉴定
engineered Saccharomyces cerevisiae
lycopene
extraction and purification
process optimization
structure identification
作者简介
石彬(1993-),男,讲师,博士,研究方向为合成生物学与发酵工程;通讯作者:邓小敏(1985-),男,副研究员,博士,研究方向为合成生物学与代谢工程。
