可口革囊星虫体腔液蛋白质的提取及其抗氧化活性研究  

Extraction and Antioxidant Properties of Protein from Phascolosoma Esculenta Coelomic Fluid

作　　者：陈纤 池海波 邱家港 陆伊俏 厉桂宁 方旭波 陈小娥 杨会成 CHEN Qian;CHI Haibo;QIU Jiagang;LU Yiqiao;LI Guining;FANG Xubo;CHEN Xiao’e;YANG Huicheng(College of Food and Pharmacy,Zhejiang Ocean University,Zhoushan 316000,China;Zhejiang International Maritime College,Zhoushan 316000,China;Zhejiang Marine Development Research Institute,Zhoushan 316021,China)

机构地区：[1]浙江海洋大学食品与药学学院,浙江舟山316000 [2]浙江国际海运职业技术学院,浙江舟山316000 [3]浙江省海洋开发研究院,浙江舟山316021

出　　处：《食品科技》2021年第7期155-160,共6页Food Science and Technology

基　　金：国家级大学生创新创业训练计划项目(201910340022);教育部产学合作研协同育人项目(201902037040,201902187008);舟山现代海洋产业创新服务综合体科技特派员专项(2020051421)。

摘　　要：文章探究了可口革囊星虫(Phascolosoma esculenta)体腔液蛋白质的抗氧化性及其与二级结构之间的关系。以可口革囊星虫体腔液为材料,用超滤分级分离,乙醇沉淀法对其蛋白质组分进行提取。通过羟基自由基清除率、总还原力及1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率等指标测定蛋白质的抗氧化性;采用傅里叶红外色谱酰胺I带光谱分峰拟合得出抗氧化蛋白质的二级结构,分析其抗氧化性和二级结构的关系。结果表明:超滤后醇沉所得分子质量<10 ku(Phascolosoma esculenta proteinⅠ,PEPⅠ)、10~30 ku(PEPⅡ)、30~50 ku(PEPⅢ)、50~100 ku(PEPⅣ)、>100 ku(PEPⅤ)5个组分粗蛋白,对羟基自由基的半数清除浓度(IC_(50))分别为3.327、1.637、5.210、6.733、0.998 mg/mL,对DPPH自由基的半数清除浓度(IC_(50))分别为12.719、5.581、9.063、11.934、2.801 mg/mL,总还原力半数清除浓度(IC_(50))分别为6.760、3.700、7.730、10.034、1.461 mg/mL。其中抗氧化性较强的2个组分,PEPⅡ二级结构的含量组成为β-折叠(40.29%)>β-转角(36.21%)>α-螺旋(23.50%)>无规卷曲(0%),PEPⅤ二级结构的含量组成为β-转角(38.28%)>α-螺旋(21.74%)>无规卷曲(20.54%)>β-折叠(19.44%)。推测β-折叠与羟基自由基清除率有较大关系,无规卷曲、β-转角与DPPH自由基清除率、总还原力有较大关系。综上所述,可口革囊星虫体腔液蛋白质有较好的抗氧化性。The antioxidant properties of coelomic fliud proteins extract from Phascolosoma esculenta and the relationship between the secondary structure were investigated.After the coelomic fluid was isolated by ultrafiltration retention separation,the protein components were extracted and separated by flocculating alcohol precipitation method.The total reducing power,hydroxyl radical scavenging rate and 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical scavenging rate were used to determine the antioxidant activity of proteins.The secondary structure of antioxidant protein was tested by Fourier infrared chromatography amideⅠband spectral peak fitting,and the relationship between the antioxidant properties and its secondary structure was analyzed.The Phascolosoma esculenta coelomic fluid protein molecular weight groups were divided into molecular weight<10 ku(Phascolosoma esculenta proteinⅠ,PEPⅠ),10~30 ku(PEPⅡ),30~50 ku(PEPⅢ),50~100 ku(PEPⅣ)and>100 ku(PEPⅤ).The half clearance concentration(IC_(50))of hydroxyl radical scavenging of PEPⅠ,PEPⅡ,PEPⅢ,PEPⅣ,PEPⅤwere 3.327,1.637,5.210,6.733,0.998 mg/mL,the IC_(50) of total reducing power of those were 6.760,3.700,7.730,10.034,1.461 mg/mL,the IC_(50) of DPPH free radical abilities of those were 12.719,5.581,9.063,11.934,2.801 mg/mL.The composition of the secondary structure content of PEPⅡwasβ-sheet(40.29%)>β-turn(36.21%)>α-helix(23.50%)>random(0%).The composition of the secondary structure content of PEPⅤwasβ-turn(38.28%)>α-helix(21.74%)>random(20.54%)>β-sheet(19.44%).It was speculated thatβ-sheet had a great relationship with hydroxyl radical scavenging,whileβ-turn and random had a great relationship with the DPPH free radical ability and the total reducing power.In general,the proteins in the coelomic fluid of Phascolosoma esculenta had better antioxidant properties.

关 键 词：可口革囊星虫 体腔液蛋白 傅里叶变换红外光谱 抗氧化性 二级结构 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]