摘要
为获得一种长心卡帕藻多糖的最佳提取工艺并测定抗过敏活性,该研究在单因素实验的基础上,以长心卡帕藻的多糖得率作为考察指标,选择料液比、超声波预处理时间、提取时间进行响应面优化实验,确定最佳工艺条件,并通过DEAE-52纤维素层析柱对提取的粗多糖进行纯化。随后利用RBL-2H3细胞模型,分别测定纯化后的长心卡帕藻多糖对RBL-2H3细胞活力、细胞脱颗粒抑制及细胞组胺释放水平调节的影响。结果表明,响应面优化后的长心卡帕藻多糖的最佳提取工艺如下:料液比(w/v)为1:92 g/mL,超声波预处理时间为40 min,提取时间为4 h,最佳提取工艺条件下,长心卡帕藻多糖提取率为13.92%。经DEAE-52纤维素层析纯化后的组分KSP(Kappaphycus alvarezii Sulfated Polysaccharides)在50~1200μg/mL浓度范围内对RBL-2H3细胞活力均未表现有显著性抑制作用,无细胞毒性;浓度为25、50及100μg/mL的KSP对经2,4-二硝基苯基白蛋白偶联物(2,4-Dinitrophenyl-Bovine Serum Albumin,DNP-BSA)刺激后的RBL-2H3细胞的β-氨基己糖苷酶酶活抑制率分别为36.3%、37.0%和51.7%,对其组胺释放抑制率分别为17.5%、18.3%和42.0%。综上,本试验得到的工艺参数具有可行性,KSP对RBL-2H3细胞无明显毒性,并表现出显著的脱颗粒和组胺的释放抑制作用,具有较好的体外抗过敏活性。该研究可为以长心卡帕藻为原料开发具有抗过敏活性的营养功能性食品提供参考。
In order to obtain the best extraction process of sulfated polysaccharides from Kappaphycus alvarezii and determine its antiallergic activity,the single factor experiments were carried out firstly.The polysaccharides yield of Kappaphycus alvarezii was chose as the investigation index,and the ratio of material to liquid,ultrasonic pretreatment time and extraction time were selected for Box-Behnken response surface design.Then the extracted crude polysaccharides was purified through the DEAE-52 cellulose chromatography column.The effects of the purified polysaccharides on RBL-2H3 cell viability,inhibition of cell degranulation and regulation of histamine release were measured by RBL-2H3 cell model method.The results showed that the optimum extraction condition of polysaccharides from Kappaphycus alvarezii was as follows:The ratio of material to liquid(w/v)was 1:92 g/mL,the ultrasonic pretreatment time was 40 min,and the extraction time was 4 h.Under the optimum extraction conditions,the extraction rate of polysaccharides from Kappaphycus alvarezii was 13.92%.The KSP(Kappaphycus alvarezii Sulfated Polysaccharides),a component purified by DEAE-52 cellulose chromatography,had no significant inhibition on the activity of RBL-2H3 cells in the concentration range from 50 to 1200μg/mL,and also had no cytotoxicity.Theβ-Hexosaminidase activity and histamine release inhibition rates of RBL-2H3 cells stimulated by 2,4-Dinitrophenyl-Bovine Serum Albumin(DNP-BSA)with 25,50 and 100μg/mL KSP were 36.3%,37.0%,51.7%,and 17.5%,18.3%,42.0%respectively.In conclusion,the process parameters obtained in this test are feasible,KSP had no obvious toxicity to RBL-2H3 cells and showed significant inhibition of degranulation and histamine release,with good in vitro anti-allergic activity.This study can provide a reference for the development of nutritional functional food with anti-allergic activity using Kappaphycus alvarezii as raw material.
作者
吴天翔
李振兴
吴燕燕
杨贤庆
李来好
陈胜军
戚勃
赵永强
WU Tianxiang;LI Zhenxing;WU Yanyan;YANG Xianqing;LI Laihao;CHEN Shengjun;QI Bo;ZHAO Yongqiang(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China;Key Laboratory of Aquatic Product Processing,Ministry of Agriculture and Rural Affairs,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;Key Laboratory of Efficient Utilization and Processing of Marine Fishery Resources of Hainan Province,Sanya Tropical Fisheries Research Institute,Sanya 572018,China)
出处
《食品工业科技》
CAS
北大核心
2023年第19期208-216,共9页
Science and Technology of Food Industry
基金
财政部和农业农村部国家现代农业产业技术体系资助(CARS-50)
中国水产科学研究院基本科研业务费资助(2020TD73)
中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助(2021SD07)。
关键词
长心卡帕藻多糖
超声波提取法
响应面优化
抗过敏活性
细胞毒性
Kappaphycus alvarezii polysaccharides
ultrasonic extraction
response surface optimization
anti-allergic activity
cytotoxicity
