超声辅助复合酶法提取小龙虾虾壳蛋白及其性质研究_汪文青.pdf

第 14 卷 第 11 期 食 品 安 全 质 量 检 测 学 报 Vol.14 No.11 2023 年 6 月 Journal of Food Safety and Quality Jun.,2023 基金项目:国家重点研发计划项目(2019YFD0902000)、湖北省农业科技创新中心项目(2022-620-000-001-25)Fund:Supported by the National Key Research and Development Program of China(2019YFD0902000),and the Hubei Agricultural Scientific and Technological Innovation Center Project(2023-620-000-001-25)*通信作者:白婵,硕士,主要研究方向为生物物理学。E-mail: 王雅,副教授,主要研究方向为功能性食品。E-mail:*Corresponding author:BAI Chan,Master,Institute of Agro-products Processing and Nuclear Agricultural Technology,Huber Academy of Agricultural Sciences,No.5 Nanhu Avenue Hongshan District,Wuhan 430064,China.E-mail: WANG Ya,Associate Professor,Lanzhou University of Technology,No.36 Pengjiaping Road,Qilihe District,Lanzhou 730050,China.E-mail: 超声辅助复合酶法提取小龙虾虾壳 蛋白及其性质研究 汪文青1,2,廖 涛2,熊光权2,王炬光2,邱 亮2,白 婵2*,王 雅1*(1.兰州理工大学生命科学与工程学院,兰州 730050;2.湖北省农业科学院农产品加工与核农技术研究所/农业农村部农产品冷链物流技术重点实验室/湖北省农产品辐照工程技术研究中心,武汉 430064)摘摘 要要:目的目的 提高副产物虾壳中蛋白的提取率,并探究虾壳蛋白的功能活性。方法方法 以小龙虾虾壳为原料,在单因素的基础上,以加酶量、复合酶比例、酶解时间和超声时间为响应变量,蛋白提取率为响应值,采用响应面 Box-Behnken 试验设计优化超声辅助复合酶法提取小龙虾壳蛋白的最佳工艺。进一步比较复合酶解与单酶酶解制备的多肽的抗氧化活性、抗血管紧张素转化酶(angiotensin converting enzyme,ACE)活性和氨基酸成分组成,研究酶解虾壳蛋白多肽的功能性质。结果结果 加酶量 10200 U,复合酶比例 0.65,酶解时间 2.8 h,超声时间 30 min,该最优条件下的蛋白质提取率为 90.30%,与预测值相对误差为 1.2%。与单酶酶解液相比,复合酶解液的抗氧化能力和 ACE 抑制能力均显著增强,清除 1,1-二苯基-2-三硝基苯肼和 2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐的半最大效应浓度分别为 4.38 mg/mL 和 3.66 mg/mL,最大的 ACE 抑制率达(44.930.89)%,且具有竞争性抑制模式。氨基酸组分分析发现酶解制备的虾壳多肽中必需氨基酸和疏水性氨基酸含量丰富,分别占氨基酸总量的 43.06%和 33.52%,从而表现出抗氧化活性和抗 ACE 活性。结论结论 该研究有效提高了虾壳蛋白的提取率,得到的虾壳多肽中富含多种必需氨基酸和疏水性氨基酸,表现出良好的抗氧化活性和 ACE 抑制活性。关键词关键词:虾壳蛋白;超声波;复合酶;抗氧化活性;抗血管紧张素转化酶抑制活性 Extraction and characterization of Procambarus clarkii shell proteins by ultrasound assisted complex enzymatic hydrolysis WANG Wen-Qing1,2,LIAO Tao2,XIONG Guang-Quan2,WANG Ju-Guang2,QIU Liang2,BAI Chan2*,WANG Ya1*(1.School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China;2.Institute of Agro-products Processing and Nuclear Agricultural Technology,Hubei Academy of Agricultural Sciences/Key Laboratory of Cold Chain Logistics Technology for Agro-product,Ministry of Agriculture and Rural Affairs/Hubei Engineering Research Center for Agricultural Products Irradiation,Wuhan 430064,China)DOI:10.19812/ki.jfsq11-5956/ts.2023.11.048第 11 期 汪文青,等:超声辅助复合酶法提取小龙虾虾壳蛋白及其性质研究 53 ABSTRACT:Objective To improve the extraction rate of protein from Procambarus clarkii shells as a by-product,and investigate the functional activity of shrimp shell proteins.Methods Using Procambarus clarkii shell as raw material,the response surface Box-Behnken test design was used to optimize the optimal extraction process of Procambarus clarkii shell protein by ultrasound assisted complex enzyme with enzyme addition amount,ratio of complex enzyme,enzymatic hydrolysis time and ultrasonic time as response variables and protein extraction rate as response values on the basis of single factor.The antioxidant activity,the angiotensin converting enzyme(ACE)inhibition ability and amino acid composition of the proteins prepared by complex enzymatic digestion and single enzyme enzymatic digestion were further compared to investigate the functional properties of the enzymatically digested Procambarus clarkii shell protein proteins.Results The protein extraction under this optimal condition was 90.30%with a relative error of 1.2%from the predicted value by adding enzyme amount of 10200 U,ratio of complex enzyme of 0.65,enzymatic digestion time of 2.8 h and ultrasound time of 30 min.Compared with the single enzyme digestion solution,the antioxidant and ACE inhibition abilities of the complex digestion solution were significantly enhanced,and the scavenging of 1,1-diphenyl-2-trinitrophenylhydrazine and 2,2-diazo-di(3-ethyl-benzothiazole-6-sulfonic acid)diamiammonium salts showed the concentration for 50%of maximal effect of 4.38 mg/mL and 3.66 mg/mL,respectively.The largest ACE suppression rate was(44.930.89)%,and there was a competitive suppression mode.The amino acid fraction analysis revealed that the enzymatically prepared Procambarus clarkii shell proteins were rich in essential and hydrophobic amino acids,accounting for 43.06%and 33.52%of the total amino acids,respectively,thus exhibiting antioxidant activity and anti-ACE activity.Conclusion proteinsThis study effectively improves the extraction rate of Procambarus clarkii shell protein,and the obtains shrimp shell proteins are rich in many essential amino acids and hydrophobic amino acids,and shows good antioxidant activity and ACE inhibitory activity.KEY WORDS:Procambarus clarkii shell proteins;ultrasound;complex enzymatic;antioxidant activity;angiotensin converting enzyme inhibitory activity 0 引 言 小龙虾,又被称为克氏原螯虾(Procambarus clarkii),蛋白质含量为 18.9%,肉质鲜美,易消化,广受消费者欢迎。2021 年,小龙虾全国产量达 263.36 万 t,加工量约为85 万 t,然而虾壳在国内小龙虾加工产业中一般按废弃物处理,虾头、虾壳等废弃物约占整虾质量的 40%左右1,这造成了巨大的资源浪费和环境污染2。因此,合理利用虾壳副产物可以提高相关产业的附加值,有较大的经济意义。小龙虾虾壳中富含蛋白质、甲壳素、虾青素、矿物质等3。其中,蛋白质约占 20%30%,其必需氨基酸模式与世界卫生组织/联合国粮食及农业组织(World Health Organization/F