富硒对黄粉虫蛋白分布及其酶解多肽抗氧化活性的影响
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摘要
本实验讨论了富硒对黄粉虫蛋白及其硒含量的影响;按照溶解性依次提取水、盐、醇和碱四种可溶性蛋白,通过原子荧光光谱仪进行硒含量的测定;对富硒黄粉虫蛋白进行酶解来获得含硒多肽,并以红细胞溶血为模型对比了富硒前后黄粉虫水解多肽的细胞抗氧化活性。结果发现,富硒后黄粉虫蛋白所占干重比例由53.86%上升至59.12%,分子量对应75 ku和37 ku的蛋白电泳条带在富硒后变深;富硒黄粉虫中(Se-TM)总硒含量为573.32μg/kg,其中有机硒含量为524.91μg/kg,占总硒含量的91.56%;在可溶性蛋白中,硒主要存在于碱溶蛋白中,占总硒的26.62%,此外,有46.63%的硒存在于不溶性组分中;富硒黄粉虫水解得到的多肽(Se-TMP)对红细胞溶血的抑制率在样品浓度为3 mg/m L时为87.32%,在实验各样品浓度下Se-TMP对红细胞氧化溶血的抑制率均高于未富硒黄粉虫水解多肽(TMP)。
The effects of selenium-enrichment on Tenebrio molitor(mealworm) protein and its selenium concentration were evaluated. Based on solubility, water-, salt-, ethanol-, and alkali-soluble proteins from T. molitor were extracted, and the selenium concentration was measured using an atomic fluorescence spectrometer. Selenium-enriched mealworm protein was hydrolyzed by enzymes to obtain Se-containing T. molitor peptide(Se-TMP), and the cellular antioxidant activities of Se-TMP and TMP were determined and compared using erythrocyte hemolysis as a model. The results showed that the dry weight ratio of mealworm proteins after Se-enrichment increased from 53.86% to 59.12% and electrophoretic bands at 75 and 37 ku were darker in Se-enriched proteins. The total selenium concentration in Se-enriched T. molitor was 573.32 μg/kg and organic selenium concentration was 524.91 μg/kg(91.56%). In soluble protein, selenium was mainly present in alkali-soluble protein, accounting for 26.62% of the total selenium. Furthermore, 46.63% of total selenium was found in the insoluble component. At a sample concentration of 3 mg/m L, the inhibition rate of Se-TMP on erythrocyte hemolysis was 87.32%. The inhibition rates of Se-TMP were all higher than those of TPM at various sample concentrations.
The effects of selenium-enrichment on Tenebrio molitor(mealworm) protein and its selenium concentration were evaluated. Based on solubility, water-, salt-, ethanol-, and alkali-soluble proteins from T. molitor were extracted, and the selenium concentration was measured using an atomic fluorescence spectrometer. Selenium-enriched mealworm protein was hydrolyzed by enzymes to obtain Se-containing T. molitor peptide(Se-TMP), and the cellular antioxidant activities of Se-TMP and TMP were determined and compared using erythrocyte hemolysis as a model. The results showed that the dry weight ratio of mealworm proteins after Se-enrichment increased from 53.86% to 59.12% and electrophoretic bands at 75 and 37 ku were darker in Se-enriched proteins. The total selenium concentration in Se-enriched T. molitor was 573.32 μg/kg and organic selenium concentration was 524.91 μg/kg(91.56%). In soluble protein, selenium was mainly present in alkali-soluble protein, accounting for 26.62% of the total selenium. Furthermore, 46.63% of total selenium was found in the insoluble component. At a sample concentration of 3 mg/m L, the inhibition rate of Se-TMP on erythrocyte hemolysis was 87.32%. The inhibition rates of Se-TMP were all higher than those of TPM at various sample concentrations.
引文
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[22]Meng-meng Zhang,Guang Wang,Hui Wu,et al.Structural characterization and immunomodulatory activity of a novel polysaccharide from Lepidiumeyenil[J].Journal of Agricultural and Food Chemistry,2016,64(9):1921-1931
[2]高柱,蔡荟梅,彭传燚,等.富硒茶叶中硒的赋存形态研究[J].中国食物与应用,2014,20(1):31-33GAO Zhu,CAI Hui-mei,PENG Chuan-yi,et al.Distribution rule and combined forms of selenium in selenium-enriched tea[J].Food and Nutrition in China,2014,20(1):31-33
[3]赵镭,杜明,张美莉,等.硒在富硒灵芝中的分布[J].中国食品学报,2005,5(4):119-123ZHAO Lei,DU Ming,ZHANG Mei-li,et al.Selenium distribution in se-enriched lucid ganoderma[J].Journal of Chinese Institute of Food Science and Technology,2005,5(4):119-123
[4]程祥,熊华,胡居吾.富硒花生油和富硒花生蛋白的基本性质研究[J].中国油脂,2013,38(7):95-97CHENG Xiang,XIONG Hua,HU Ju-wu.Basic properties of selenium enriched peanut oil and protein[J].China Oils and Fats,2013,38(7):95-97
[5]余芳,汪社英,方勇,等.富硒绿茶硒蛋白的提取工艺研究[J].南京农业大学学报,2008,31(4):140-143YU Fang,WANG She-ying,FANG Yong,et al.Study on extraction of selenium-containing protein from selenium-enriched green tea[J].Journal of Nanjing Agricultural University,2008,31(4):140-143
[6]张建新,孙长江.黄粉虫抗菌肽的分离纯化及生物活性研究[J].现代食品科技,2016,32(1):116-121ZHANG Jian-xin,SUN Chang-jiang.Separation,purification and biological activity evaluation of antimicrobial peptides from tenebrio molitor[J].Modern Food Science and Technology,2016,32(1):116-121
[7]张莉莎,张建新,欧晓锋,等.黄粉虫抗氧化活性肽的分离纯化研究[J].食品科学,2009,30(22):180-183ZHANG Li-sha,ZHANG Jian-xin,OU Xiao-feng.Separation and purification of antioxidant peptides from yellow mealworm[J].Food Science,2009,30(22):180-183
[8]代春华.黄粉虫幼虫蛋白酶解制备降血压肽的研究[D].无锡:江南大学,2012DAI Chun-hua.Study on antihypertensive peptides from Tenebrio molitor larva protein htdrolysate[D].Wuxi:Jiangnan University,2012
[9]Sunde R A.Selenium.In:Ross AC,Caballero B,Cousins RJ,Tucker KL,Ziegler TR,eds.Modern nutrition in health and disease.11th ed[M].Philadelphia,PA:Lippincott Williams&Wilkins,2012
[10]Sunde R A.Selenium.In:Bowman B,Russell R,eds.Present knowledge in nutrition.9th ed[M].Washington,D C:International Life Sciences Institute,2006
[11]Terry E N,Diamond A M.Selenium.In:Erdman JW,Macdonald IA,Zeisel SH,eds.Present knowledge in nutrition.10th ed[M].Washington,DC:Wiley-Blackwell,2012
[12]Sunde R A.Selenium.In:Coates PM,Betz JM,Blackman MR,et al.Encyclopedia of dietary supplements.2nd ed[M].London and New York:Informa Healthcare,2010
[13]Institute of Medicine,Food and Nutrition Board.Dietary reference intakes:vitamin c,vitamin e,selenium,and carotenoids[M].Washington,DC:National Academy Press,2000
[14]Rayman M P.Food-chain selenium and human health:emphasis on intake[J].Br.J.Nutr.,2008,100(2):254-68
[15]Rayman M P.Selenium and human health[J].Lancet,2012,379:1256-68
[16]Verena Nowak,Diedelinde Persijn,Doris Rittenschober,et al.Review of food composition data for edible insects[J].Food Chemistry,2016,193:39-46
[17]柯根林J E.精编蛋白质科学实验指南[M].北京:科学出版社,2007Ke Genlin J E.Guide to refined protein science experiment[M].Beijing:Science Press,2007
[18]张虹,张猛猛,赖富饶,等.甜菜碱保护细胞免受AAPH损伤的研究[J].现代食品科技,2016,32(6):18-24ZHANG Hong,ZHANG Meng-meng,LAI Fu-rao,et al.Protective effects of betaine on 2,2-Azobis(2-amidinopropane)dihydrochloride(AAPH)-induced oxidative stress in cells[J].Modern Food Science and Technology,2016,32(6):18-24
[19]Clemens M R,Waller H D.Lipid peroxidation in erythrocytes[J].Chemistry and Physics of Lipids,1987,45:251-268
[20]Sarah van Broekhoven,Shanna Bastiaan-Net,Nicolette WDe Jong,et al.Influence of processing and in vitro digestion on the allergic cross-reactivity of three mealworm species[J].Food Chemistry,2016:1075-1083
[21]任尧.宽体金线蛭/东亚钳蝎分离蛋白中抗凝血、抗疲劳活性肽的酶法制备及其结构鉴定研究[D].广州:华南理工大学,2015REN Yao.Study on enzymatic preparation and identification of anticoagulant peptides from whitmaniapigra and buthusmartensii kirsch protein isolation hydrolysates and their antifatigue effect in vivo[D].Guangzhou:South China University of Technology,2015
[22]Meng-meng Zhang,Guang Wang,Hui Wu,et al.Structural characterization and immunomodulatory activity of a novel polysaccharide from Lepidiumeyenil[J].Journal of Agricultural and Food Chemistry,2016,64(9):1921-1931