食品与发酵工业 >

2020 , Vol. 46 >Issue 23: 21 - 28

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.024605

研究报告

皱纹盘鲍(Haliotis discus hannai Ino)肌肉酶解产物制备及其免疫活性研究

  • 任鼎鼎 ,
  • 黄梦怡 ,
  • 郑惠娜 ,
  • 曹文红 ,
  • 林海生 ,
  • 秦小明 ,
  • 章超桦
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  • 1(广东海洋大学 食品科技学院,广东 湛江,524088);
    2(广东海洋大学 深圳研究院,广东 深圳,518108);
    3(国家贝类加工技术研发分中心(湛江),广东省水产品加工与安全重点实验室,广东省海洋生物制品工程实验室,广东 湛江,524088);
    4(海洋食品精深加工关键技术省部共建协同创新中心(大连工业大学),辽宁 大连,116034)
硕士研究生(郑惠娜副教授为通讯作者,E-mail:zhenghn@gdou.edu.cn)

收稿日期: 2020-06-01

  修回日期: 2020-07-09

  网络出版日期: 2020-12-30

基金资助

广东省自然科学基金项目(2016A030313810);广东省现代农业产业技术体系创新团队项目(2019KJ146);国家贝类产业技术体系项目(CARS-49)

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Preparation and immunoactivity of enzymatically hydrolyzed products of abalone (Haliotis discus hannaiIno) muscleREN

  • Dingding ,
  • HUANG Mengyi ,
  • ZHENG Huina ,
  • CAO Wenhong ,
  • LIN Haisheng ,
  • QIN Xiaoming ,
  • ZHANG Chaohua
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  • 1(College of Food Science and Technology,Guangdong Ocean University,Zhanjiang 524088,China);
    2(Shenzhen Institute of Guangdong Ocean University,Shenzhen 518108,China);
    3(National Research and Development Branch Center for Shellfish Processing (Zhanjiang),Guangdong Provincial Key Laboratory of Aquatic Products Processing and Safety,Guangdong Province Engineering Laboratory for Marine Biological Products,Zhanjiang 524088,China);
    4(Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,China)

Received date: 2020-06-01

  Revised date: 2020-07-09

  Online published: 2020-12-30

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摘要

为开发鲍鱼生物制品,对鲍鱼肌肉酶解产物的制备以及免疫活性进行研究。以皱纹盘鲍(Haliotis discus hannai Ino)肌肉为原料,采用动物蛋白酶和风味蛋白酶进行酶解,同时以蛋白回收率和水解度为指标优化酶解工艺条件,获得鲍鱼酶解多肽(abalone enzymatic hydrolysis of peptides,AHP),进一步采用斑马鱼动物实验模型,对AHP免疫活性进行初步评价分析。结果表明,酶解4 h时,AHP的蛋白回收率和水解度最佳,分别为(55.88±0.03)%和(20.32±0.01)%。体内免疫活性实验结果显示:各样品均能够提高斑马鱼模型的溶菌酶(lysozyme,LYS)活性、碱性磷酸酶(alkaline phosphatase,ALP)活性、免疫球蛋白G(immunoglobulinG,IgG)含量,其中,喂养冷冻干燥样品组中LYS活性最高达到(166.04±23.48) U/L、ALP活性最高达到(7.46±1.02) IU/L、IgG含量最高达到(0.31±0.09) μg/mL,与空白对照组相比具有显著性差异(P＜0.05);在超滤组分中,与＞10 kDa和5 k~10 kDa样品组相比,＜5 kDa超滤样品组的活性明显较高(P＜0.05),LYS活性最高为(1481.29±160.70) U/L、ALP活性最高为(1617.39±206.70) IU/L、IgG含量最高为(30.74±1.73)μg/mL。结果显示,鲍鱼酶解多肽具有一定的免疫活性,并且＜5 kDa超滤组分免疫活性较强。

关键词： 鲍鱼肌肉; 酶解产物; 超滤组分; 斑马鱼模型; 免疫活性

本文引用格式

任鼎鼎 , 黄梦怡 , 郑惠娜 , 曹文红 , 林海生 , 秦小明 , 章超桦 . 皱纹盘鲍(Haliotis discus hannai Ino)肌肉酶解产物制备及其免疫活性研究[J]. 食品与发酵工业, 2020 , 46(23) : 21 -28 . DOI: 10.13995/j.cnki.11-1802/ts.024605

Abstract

In order to develop bioproducts from abalone (Haliotis discus hannaiIno),abalone muscle was enzymatically hydrolyzed and the immunoactivity of the hydrolysate was analyzed.Animal protease and flavor protease preparations were used for enzymolysis of abalone muscle,and protein recovery rate and degree of hydrolysis (DH) were used as indexes for optimization of the enzymolysis conditions.The immunoactivity of enzymatically hydrolyzed peptides (AHP) was analyzed with zebrafish model.The results showed that optimal protein recovery and DH were obtained after 4h hydrol-ysis,which were (55.88±0.03)% and (20.32±0.01)%,respectively.The results ofin vitro immunoactivity experiments showed that AHP increased the activities of lysozyme (LYS) and alkaline phosphatase (ALP) and level of IgG in zebrafish model,of which the activity of LYS and ALP reached (166.04±23.48) U/L and (7.46±1.02) IU/L,respectively,and the IgG reached (0.31±0.09) μg/mL in the freeze-dried sample group,which were significantly different from that of the control group (P<0.05).Among the ultrafiltration components,compared with the >10 kDa and 5 k-10 kDa sample groups,the activity of the <5 kDa ultrafiltration sample group was significantly higher (P<0.05),the highest activity of LYS and ALP were (1481.29±160.70) U/L and (1 617.39±206.70) IU/L,respectively,and IgG was up to (34.77±2.40) μg/mL.AHP exhibited certain immunoactivity and the <5 kDa group ultrafiltration components were the most effective.

Key words： abalone muscle; enzymatically hydrolyzed products; ultrafiltration components; zebrafish model; immunoactivity

参考文献

[1] 张孙现.不同干燥方法对鲍鱼营养成分的影响[J].食品科学技术学报,2018,36(2):78-83.
ZHANG S X.Effect of drying methods to nutritional compositions of abalone[J].Journal of Food Science and Technology,2018,36(2):78-83.
[2] 林位琅,林光文,黄洪龙,等.2018年鲍养殖分析[J].科学养鱼,2019(6):1-3.
LIN W L,LIN G W,HUANG H L,et al.Analysis on abalone culture in 2018[J].Scientific Fish Farming,2019(6):1-3.
[3] 农业部渔业渔政管理局.中国渔业统计年鉴[M].北京:中国农业出版社,2019.
Fisheries and Fisheries Administration of the Ministry of Agriculture.China fishery statistical yearbook[M].Beijing:China Agriculture Press,2019.
[4] 刘先进,陈胜军,李来好,等.四种鲍鱼肌肉营养成分分析与品质评价[J].食品与发酵工业,2018,44(5):227-231.
LIU X J,CHEN S J,LI L H,et al.Nutritional analysis and quality evaluation of four kinds of abalone muscle[J].Food and Fermentation Industries,2018,44(5):227-231.
[5] 张其标,孙梦莹,郝林娟,等.鲍鱼腹足肌肉成分和组织微观结构的季节差异研究[J].食品工业科技,2020,41(5):7-11.
ZHANG Q B,SUN M Y,HAO L J,et al.Study on seasonal difference of composition and micro-structure of abalone(Haliotis discus hannaiIno) gastropod muscle[J].Food Industry Technology,2020,41(5):7-11.
[6] 高智春.鲍鱼干制工艺及产品特性的研究[D].福州:福建农林大学,2011.
GAO Z C.Study on drying technology and product characteristics of abalone[D].Fuzhou:Fujian Agriculture and Forestry University,2011.
[7] 朱蓓薇,吴厚刚,董秀萍,等.鲍鱼软包装即食产品的制备方法:中国,CN101019664A[P].2007-08-22.
ZHU B W,WU H G,DONG X P,et al.Preparation method of abalone soft-packaged instant product:China,CN101019664A[P].2007-08-22.
[8] 张帅.河蚬酶解产物解酒护肝活性的研究[D].湛江:广东海洋大学,2019.
ZHANG S.Study on the anti-alcohol and liver-protection efficacy of hydrolysate from soft tissue ofCorbicula fluminea[D].Zhanjiang:Guangdong Ocean University,2019.
[9] LEE S H,QIAN Z J,KIM S K.A novel angiotensin I converting enzyme inhibitory peptide from tuna frame protein hydrolysate and its antihypertensive effect in spontaneously hypertensive rats[J].Food Chemistry,2010,118(1):96-102.
[10] JANG H L,LICEAGA A M,YOON K Y.Purification,characterisation and stability of an antioxidant peptide derived from sandfish(Arctoscopus japonicus) protein hydrolysates[J].Journal of Functional Foods,2016,20:433-442.
[11] LI W,XU C,ZHANG C H,et al.The purification and identification of immunoregulatory peptides from oyster(Crassostrea hongkongensis) enzymatic hydrolysate[J].RSC Advances,2019,9(56):32 854-32 863.
[12] ZHOU D Y,TANG Y,ZHU B W,et al.Antioxidant activity of hydrolysates obtained from scallop (Patinopecten yessoensis) and abalone(Haliotis discus hannai Ino) muscle[J].Food Chemistry,2012,132(2):815-822.
[13] 王军玲,魏配晓,邱绪建,等.鲍鱼肌肉多糖的性质及抗肿瘤活性[J].食品科学,2018,39(12):26-32.
WANG J L,WEI P X,QIU X J,et al.Physicochemical properties and antitumor activity of polysaccharides from abalone muscle[J].Food Science,2018,39(12):26-32.
[14] LIU X B,HARADA N,YAMANE S,et al.Effects of long-term dipeptidyl peptidase-IV inhibition on body composition and glucose tolerance in high fat diet-fed mice[J].Life Sciences,2009,84(25):876-881.
[15] GONÇALVES A,MARQUES C,LEAL E,et al.Dipeptidyl peptidase-IV inhibition prevents blood-retinal barrier breakdown,inflammation and neuronal cell death in the retina of type 1 diabetic rats[J].Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease,2014,1842(9):1 454-1 463.
[16] 吉薇,章超桦,ALLAN V K,等.糖尿病斑马鱼模型的建立与南极磷虾酶解物降血糖活性评价[J].食品与机械,2019,35(6):24-29.
JI W,ZHANG C H,ALLAN V K,et al.Establishment of diabetic zebrafish model and evaluation of hypoglycemic activity of antarctic krill enzymatic hydrolysates[J].Food and Machinery,2019,35(6):24-29.
[17] MICHELLE D,RACHEL E H,LEVI A G,et al.Long-term drug administration in the adult zebrafish using oral gavage for cancer preclinical studies[J].Disease Models and Mechanisms,2016,9(7):811-820.
[18] DINDAY M T,BARABAN S C.Large-scale phenotype-based antiepic-leptic drug screening in a zebrafish model of dravet synodrome(1,2,3)[J].Eneuro,2015,2(4):1-19.
[19] WANG G L,RAJPUROHIT S K,DELASPRE F,et al.First quantitative high-throughput screen in zebrafish identifies novel pathway for increasing pancreatic β-cell mass[J].Elife,2015,4:1-19.
[20] 刘发生,王荣春,郭敬兰,等.利用斑马鱼模型研究芦笋有效成分对免疫功能的调节作用[J].山东科学,2017,30(6):29-34.
LIU F S,WANG R C,GUO J L,et al.The immune-regulatory effects of active ingredients of asparagus using zebrafish model[J].Shandong Science,2017,30(6):29-34.
[21] 李婉.牡蛎免疫活性成分的分离纯化及产品开发[D].湛江:广东海洋大学,2017.
LI W.Separation and purification of oyster immune active ingredients and product development[D].Zhanjiang:Guangdong Ocean University,2017.
[22] 中国国家标准化管理委员会.GB/T 5009.3—2016 食品中水分的测定方法[S].北京:中国标准出版社,2016.
China National Standardization Management Committee.GB/T 5009.3—2016 Method for determination of moisture in food[S].Beijing:China Standards Press,2016.
[23] 中国国家标准化管理委员会.GB/T 5009.6—2016 食品中脂肪的测定方法[S].北京:中国标准出版社,2016.
China National Standardization Management Committee.GB/T 5009.6—2016 Method for determination of fat in food[S].Beijing:China Standards Press,2016.
[24] 中国国家标准化管理委员会.GB/T 5009.4—2016 食品中灰分的测定方法[S].北京:中国标准出版社,2016.
China National Standardization Management Committee.GB/T 5009.4—2016 Method for determination of ash in food[S].Beijing:China Standards Press,2016.
[25] 中国国家标准化管理委员会.GB/T 9695.31—2008 肉制品总糖的测定方法[S].北京:中国标准出版社,2008.
China National Standardization Management Committee.GB/T 9695.31—2008.Method for determination of total sugar in meat products[S].Beijing:China Standards Press,2008.
[26] 中国国家标准化管理委员会.GB/T 5009.5—2016 食品中蛋白质的测定方法[S].北京:中国标准出版社,2016.
China National Standardization Management Committee.GB/T 5009.5—2016 Method for determination of protein in food[S].Beijing:China Standards Press,2016.
[27] 王永华.食品分析[M].北京:中国轻工业出版社,2012.
WANG Y H.Food analysis[M].Beijing:China Light Industry Press,2012.
[28] YOUSEFI S,HOSEINIFAR S H,HAMED P,et al.The effects of dietary supplement of galactooligosaccharide on innate immunity,immune related genes expression and growth performance in zebrafish(Danio rerio)[J].Fish &Shellfish Immunology,2018,73:192-196.
[29] SHEIKHZADEH N,PASHAKI A K,NOFOUZI K,et al.Effects of dietary Ergosan on cutaneous mucosal immune response in rainbow trout (Oncorhynchus mykiss)[J].Fish &Shellfish Immunology,2012,32(3):407-410.
[30] SHEIKHZADEH N,KOUCHAKI M,MEHREGAN M,et al.Influence of nanochitosan/zeolite composite on growth performance,digestive enzymes and serum biochemical parameters in rainbow trout (Oncorhynchus mykiss)[J].Aquaculture Research,2017,48:5 955-5 964.
[31] 张华丹,张玲云,张国玉,等.响应面法优化南极磷虾蛋白酶解工艺条件[J].食品工业,2019,40(7):94-98.
ZHANG H D,ZHANG L Y,ZHANG G Y,et al.Optimization of proteolytic conditions of antarctic krill by response surface methodology[J].Food Industry,2019,40(7):94-98.
[32] 柏昌旺,章超桦,林海生,等.响应面法优化制备牡蛎短肽工艺[J].广东海洋大学学报,2019,39(6):85-92.
BAI C W,ZHANG C H,LIN H S,et al.Optimization of preparation process of oysteroligopeptides by response surface methodology[J].Journal of Guangdong Ocean University,2019,39(6):85-92.
[33] XIA I F,CHEUNG J S,WU M,et al.Dietary chitosan-selenium nanoparticle(CTS-SeNP) enhance immunity and disease resistance in zebrafish[J].Fish &Shellfish Immunology,2019,87:449-459.
[34] 王裕玉,周歧存,卜宪勇,等.饲料中不同含量豆粕对乌苏里拟鲿生长、体成分及表观消化率的影响[J].水生生物学报,2016,40(4):681-689.
WANG Y Y,ZHOU Q C,BU X Y,et al.Effects of replacing fish meal with soybean meal on growth performance,body composition and apparent digestibility coefficients ofPseudobagrus ussuriensis[J].Journal of Hydrobiology,2016,40(4):681-689.
[35] MERCIER A,GAUTHIER S F,FLISS I.Immunomodulating effects of whey proteins and their enzymatic digests[J].International Dairy Journal,2004,14(3):175-183.
[36] FENG L,HE W,JIANG J,et al.Effects of dietary pyridoxine on disease resistance,immune responses and intestinal microflora in juvenile Jian carp (Cyprinus carpiovar.Jian)[J].Aquaculture Nutrituon,2010,16(3):254-261.
[37] YANG Y,PARK J,YOU S G,et al.Immuno-stimulatory effects of sulfated polysaccharides isolated fromCodium fragile in olive flounder,Paralichthys olivaceus[J].Fish &Shellfish Immunology,2019,87:609-614.
[38] 艾春香,陈立侨,温小波,等.VE对河蟹血清和组织中超氧化物歧化酶及磷酸酶活性的影响[J].台湾海峡,2003(1):24-31.
AI C X,CHEN L Q,WEN X B,et al.Effect of vitamin E on SOD,ALP and ACP activities of Chinese mitten-handed crab,Eriocheir sinensis[J].Taiwan Straits,2003(1):24-31.
[39] 江贤敏,王正明,潘利华,等.提取温度对霍山石斛多糖理化性质及肠道黏膜免疫活性的影响[J].食品科学,2017,38(7):176-181.
JIANG X M,WANG Z M,PAN L H,et al.Effect of extraction temperature on physicochemical properties and intestinal mucosal immunomodulating activity of polysaccharides fromDendrobium huoshanense[J].Food Science,2017,38(7):176-181.
[40] 柳旭东,梁萌青,张利民,等.饲料中添加水解鱼蛋白对半滑舌鳎稚鱼生长及生理生化指标的影响[J].水生生物学报,2010,34(2):242-249.
LIU X D,LIANG M Q,ZHANG L M,et al.Effect of fish protein hydrolysate levels on growth performance and biological and physiological parameters in tongue sole(Cynoglossus semilaevis Günther,1873)post-larvae[J].Journal of Hydrobiology,2010,34(2):242-249.
[41] 陈慕雁,张秀梅,连建华.大菱鲆仔稚鱼期消化酶及碱性磷酸酶活性的变化[J].中国海洋大学学报:自然科学版,2005(3):140-143.
CHEN M Y,ZHANG X M,LIAN J H.Development of some digestive enzymes and alkaline phosphatase activities in turbotScophthalmus maximus larvae and juveniles[J].Journal of Ocean University of China:Natural Science Edition,2005(3):140-143.
[42] ZHANG W N,ZHANG M X,CHENG A Y,et al.Immunomodulatory and antioxidant effects ofAstragalus polysaccharide liposome in large yellow croaker (Larimichthys crocea)[J].Fish &Shellfish Immunology,2020,100:126-136.
[43] 万发武,熊洁.免疫球蛋白治疗小儿川崎病的应用与合适剂量分析[J].首都食品与医药,2019,26(24):64-65.
WAN F W,XIONG J.The application of immunoglobulin in the treatment of children’s Kawasaki disease and the analysis of appropriate dose[J].Capital Food and Medicine,2019,26(24):64-65.
[44] 温世芳.人免疫球蛋白联合重组人干扰素α2b注射液治疗重症手足口病患儿的临床疗效观察[J].首都食品与医药,2020,27(1):73.
WEN S F.Observation on the clinical efficacy of human immunoglobulin combined with recombinant human interferon α2b injection in the treatment of children with severe Hand,Foot and Mouth disease[J].Capital Food and Medicine,2020,27(1):73.
[45] LIU J Y,WEI T Z,WU X,et al.Early exposure to environmental levels of sulfamethoxazole triggers immune and inflammatory response of healthy zebrafish larvae[J].Science of The Total Environment,2020,703:134 724.
[46] 公绪鹏,李宝山,张利民,等.饲料蛋白质和能量含量对云纹龙胆石斑鱼幼鱼生长、体组成及消化酶活力的影响[J].渔业科学进展,2018,39(2):85-95.
GONG X P,LI B S,ZHANG L M,et al.Effects of dietary protein and energy levels on growth,body composition and digestive enzymes activities of juvenile hybrid grouper,Epinephelus lanceolatus×E.moara[J].Fishery Science Progress,2018,39(2):85-95.
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