方格星虫酶解工艺优化及酶解物免疫活性  

Optimization of Enzymatic Hydrolysis of Sipunculus nudus L. and Immunoassay of Its Enzymatic Hydrolysis

作　　者：孙瑞坤[1] 章超桦[1,2,3,4] 曾少葵 秦小明[1,2,3,4] 曹文红 高加龙[1,2,3,4] 郑惠娜 SUN Rui-kun;ZHANG Chao-hua;ZENG Shao-kui;QIN Xiao-ming;CAO Wen-hong;GAO Jia-long;ZHENG Hui-na(College of Food Science and Technology,Guangdong Ocean University;Guangdong Provincial Key Laboratory of Aquatic Products Processing and Safety;Key Laboratory of Advanced Processing of Aquatic Products of Guangdong Higher Education Institution;National Research and Development Branch Center for Shellfish Processing.Zhanjiang 520488,China)

机构地区：[1]广东海洋大学食品科技学院 [2]广东省水产品加工与安全重点实验室 [3]广东普通高等学校水产品深加工重点实验室 [4]国家贝类加工技术研发分中心(湛江),广东湛江524088

出　　处：《广东海洋大学学报》2018年第3期54-61,共8页Journal of Guangdong Ocean University

基　　金：广东省教育厅创新强校工程项目(2014GKXM047);广东普通高等学校水产品高值化加工与利用创新团队项目(GDOU2016030503);2016年广东省应用型科技研发专项资金项目(2016B020235002)

摘　　要：【目的】优化方格星虫(Sipunculus nudus L.)酶解工艺条件,探讨其酶解液对细胞免疫活性的影响。【方法】在确定实验用酶和单因素实验基础上,利用响应面设计建立数学模型,以水解度为响应值,进行3因素3水平的响应面分析,得到最适酶解条件。以小鼠脾淋巴细胞的增殖、小鼠腹腔巨噬细胞生成NO量和吞噬能力来评价酶解液对免疫细胞的促进作用。【结果】以动物水解蛋白酶和风味蛋白酶组合用酶效果最佳,方格星虫酶解的适宜条件为pH 7.0、加酶量5 000 U/g、液料比3∶1,55℃酶解5.2 h,其水解度为38.92%;以此条件得到的酶解液进行实验,酶解液质量浓度为0.9 mol/mL时,小鼠脾淋巴细胞的增殖率为30.57%,小鼠腹腔巨噬细胞生成NO能力为56.2μmol/L,吞噬能力D(540 nm)值均明显高于阳性对照组(P<0.01)。【结论】方格星虫酶解液对细胞免疫具有促进作用。【Objective】To Optimize the enzymolysis process conditions of Sipunculus nudus L. and explore the effect of enzymatic hydrolysates on cellular immune activity.【Method】Based on the determination of experimental enzymes and single factor experiments, the response surface design was used to establish a mathematical model. With the degree of hydrolysis as the response value, response surface analyses of 3 factors and 3 levels were performed to obtain the optimal conditions for enzymatic hydrolysis. The proliferation of mouse spleen lymphocytes, NO production in peritoneal macrophages of mice and phagocytosis capacity were used to evaluate the effect of the enzymatic hydrolysate on immune cells.【Result】Animal proteases and flavour proteases were used as experimental enzymes. The optimum conditions for the enzymatic hydrolysis of S. nudus L. were pH 7.0, enzyme addition amount 5 000 U/g, ratio of liquid to solid 3∶1, and hydrolysis at 55 ℃ for 5.2 h. The degree of hydrolysis was 38.92%. The enzymatic hydrolysate obtained under these conditions was tested at a concentration of 0.9 mol/mL, the proliferation rate of mouse spleen lymphocytes was 30.57%, and the ability of mouse peritoneal macrophages to generate NO was 56.2 μmol/L. The OD540 nm value was significantly higher than those in the positive control group（P〈0.01）.【Conclusion】The enzymatic hydrolysate of S. nudus L. has a promoting effect on cellular immunity.

关 键 词：方格星虫 酶解 响应面分析 免疫调节 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]