墨鱼缠卵腺糖蛋白提取工艺优化及纯化
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摘要
本文以墨鱼缠卵腺为实验原料,通过单因素实验及响应面试验优化墨鱼缠卵腺糖蛋白的提取工艺,随后对最优工艺下得到的糖蛋白进行分离纯化,电泳分析其纯度,并进一步研究其蛋白及总糖含量。结果表明,墨鱼缠卵腺糖蛋白提取最佳工艺为提取温度24℃,提取时间3.6 h,NaOH浓度0.4 mol/L,料液比1∶20 g/mL,超声功率200 W。在此条件下进行验证试验,得出墨鱼缠卵腺糖蛋白实际得率为16.13%±0.16%;经DEAE-52纤维素离子交换柱层析、Sephacryl S-300HR柱层析分离后得到纯品糖蛋白,电泳分析表明所纯化的糖蛋白纯度达到电泳纯,其中蛋白含量为65.53%±0.8%,糖含量为31.74%±1.1%,得率为墨鱼缠卵腺去外层薄膜原料的0.91%。
The nidamental gland of cuttlefish was used as experimental material,and the single factor experiments and response surface test were used to optimize the extraction process of nidamental gland glycoprotein from cuttlefish.Then the glycoprotein obtained under that was separated and purified. And its purity was analyzed by electrophoresis,its protein and toted sugar content were follow researched. The results showed that the best extraction process of nidamental gland glycoprotein was:extraction temperature 24 ℃,extraction time 3.6 h,the Na OH concentration 0.4 mol/L,the ratio of material to liquid 1∶ 20 g/mL and the ultrasonic power 200 W.Under the condition of the verification tests,the actual yield of nidamental gland glycoprotein was 16.13% ± 0.16%. Purified by DEAE-52 celloulus ion exchange column chromatography and Sephacryl S-300 HR column chromatography was to obtain pure glycoprotein.The electrophoresis analysis showed that the purity of the purified glycoprotein reached electrophoresis purity,and the protein content was 65.53% ± 0.8%,and the sugar contents was 31.74% ± 1.1%. The yield was 0.91% of the nidamental gland to the outer film material.
The nidamental gland of cuttlefish was used as experimental material,and the single factor experiments and response surface test were used to optimize the extraction process of nidamental gland glycoprotein from cuttlefish.Then the glycoprotein obtained under that was separated and purified. And its purity was analyzed by electrophoresis,its protein and toted sugar content were follow researched. The results showed that the best extraction process of nidamental gland glycoprotein was:extraction temperature 24 ℃,extraction time 3.6 h,the Na OH concentration 0.4 mol/L,the ratio of material to liquid 1∶ 20 g/mL and the ultrasonic power 200 W.Under the condition of the verification tests,the actual yield of nidamental gland glycoprotein was 16.13% ± 0.16%. Purified by DEAE-52 celloulus ion exchange column chromatography and Sephacryl S-300 HR column chromatography was to obtain pure glycoprotein.The electrophoresis analysis showed that the purity of the purified glycoprotein reached electrophoresis purity,and the protein content was 65.53% ± 0.8%,and the sugar contents was 31.74% ± 1.1%. The yield was 0.91% of the nidamental gland to the outer film material.
引文
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[21]周国杰,黄晓玲,范秀萍.盐提法提取波纹巴非蛤全脏器中糖蛋白的工艺条件研究[J].现代农业科技,2009,21(1):260-261.
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[24]刘晓飞,刘宁,张娜.超声波辅助酶法提取甜玉米穗轴黄酮及抑菌性检测[J].食品科学,2014,35(20):79-82.
[25]Zou D,Yu F U,Chen S,et al.Optimization of the extraction technology of HERBA CENTIPEDAE by central composite design-response surface method[J].Medicinal Plant,2015,6(29):21-24.
[26]张妍,高蕾,王正红,等.响应面实验优化喷雾干燥制备核桃分心木速溶粉及其冲调性分析[J].食品科学,2016,37(18):47-51.
[27]Shivanna N,Naika M,Khanum F.Antioxidant,antidiabetic and renal protective properties of Stevia rebaudiana[J].Journal o Diabetes and Its Complications,2013,27(2):103-113.
[2]宋超霞,王春琳,邵银文,等.野生与养殖曼氏无针乌贼肌肉的营养成分和评价[J].营养学报,2009,31(3):301-303.
[3]侯清娥,董建伟,王录军.糖蛋白的研究概述[J].农产品加工,2016,1(1):69-71.
[4]谭艾娟,梁宗琦,刘爱英.蝉拟青霉菌丝体糖蛋白的分离提取及组分鉴定[J].食品科学,2007,28(1):159-160.
[5]包郁明,陶宇,候虎,等.鲍鱼脏器糖蛋白超声波辅助提取工艺及分离纯化的研究[J].食品与发酵工业,2011,31(9):11-14.
[6]王倩,刘淑集,林彩平,等.响应面法优化鱿鱼缠卵腺糖蛋白提取工艺研究[J].食品工业科技,2014,35(3):261-265.
[7]任国艳,李八方,赵雪,等.海蜇头糖蛋白超声辅助提取工艺研究[J].农业工程学报,2008(2):255-259.
[8]Kisugi J,Ohye H,Kamiya H,et al.Biopolymers from marine invertebrates X,mode of action of an antibacterial glycoprotein,aplysianin E,from eggs of a sea here Aplysia kurodai[J].Chemical and Pharmaceutical Bulletin,1989,37(11):3050-3053.
[9]曾婷婷,徐明生,蒋艳.超声波法提取河蚬糖蛋白[J].江西农业大学学报,2008,30(1):144-148.
[10]Shigeru K,Yoko S,Haruo M,et al.Occurrence of a mucintype glycoprotein in nidamental gland mucosubstance from the squid Illex argentinus[J].Fisheries Science,2008,60(2):193-197.
[11]Lei X L,Fan X P,Zhao S J,et al.Study on extracting and fractionating of glycoprotein of dried octopus[J].Journal of Shaanxi University of Sclence and Technolog,2006,24(4):45-49.
[12]Su Y T,Xu Y J.Study on the extraction and purification of glycoprotein from the yellow seahorse,Hippocampus kuda Bleeker[J].Food Science and Nutrition,2015,3(4):302-312.
[13]Dai H J,Sun Y L,Zheng X L,et al.Extraction optimization,preliminary characterization and antioxidant activity of glycoproteins from the muscle of Sepia pharaonis[J].Food Science and Technology Research,2016,22(1):39-52.
[14]Jiao B,Cassano A,Drioli E.Recent advances on membrane processes for the concentration of fruit juices:A review[J].Journal of Food Engineering,2004,63:303-324.
[15]程玉祥.茶树叶糖蛋白的纯化研究[D].合肥:安徽农业大学,2002.
[16]Wang Y,Zou T T,Xiang M H,et al.Purification and characterization of a soluble glycoprotein from garlic(Allium sativum)and its in vitro bioactivity[J].Preparative Biochemistry and Biotechnology,2016,46(7):709-716.
[17]王倩.太平洋褶柔鱼缠卵腺糖蛋白的分离提取及活性分析[D].福州:福建农林大学,2013.
[18]Senthilkumar D,Jayanthi S.Partial characterization and anticancer activities of purified glycoprotein extracted from green seaweed Codium decorticatum[J].Journal of Functional Foods,2016,25:323-332.
[19]韦一能.竹荪中糖蛋白的聚丙烯酸胺凝胶电泳分析[J].广西师范大学学报:自然科学版,1991,9(1):57-58.
[20]方旭波,陈小娥,余辉,等.鱿鱼硫酸软骨素糖蛋白的分离纯化和鉴定[J].中国食品学报,2009,9(4):103-109.
[21]周国杰,黄晓玲,范秀萍.盐提法提取波纹巴非蛤全脏器中糖蛋白的工艺条件研究[J].现代农业科技,2009,21(1):260-261.
[22]中华人民共和国卫生部.GB/T14772-2008食品中粗脂肪的测定[S].北京:中国标准出版社,2008.
[23]焦庆才.糖工程概论[M].北京:科学出版社,2010:45.
[24]刘晓飞,刘宁,张娜.超声波辅助酶法提取甜玉米穗轴黄酮及抑菌性检测[J].食品科学,2014,35(20):79-82.
[25]Zou D,Yu F U,Chen S,et al.Optimization of the extraction technology of HERBA CENTIPEDAE by central composite design-response surface method[J].Medicinal Plant,2015,6(29):21-24.
[26]张妍,高蕾,王正红,等.响应面实验优化喷雾干燥制备核桃分心木速溶粉及其冲调性分析[J].食品科学,2016,37(18):47-51.
[27]Shivanna N,Naika M,Khanum F.Antioxidant,antidiabetic and renal protective properties of Stevia rebaudiana[J].Journal o Diabetes and Its Complications,2013,27(2):103-113.