吴羽茜,王瑛,林晓姿,陈秉彦,何志刚

• 1:福建省农业科学院农产品加工研究所
• 2:福建省农产品(食品)加工重点实验室

摘要(Abstract)：

为了明确金枪鱼蒸煮液蛋白酶解物的理化特征，并探究其促成骨细胞增殖的活性，采用6种蛋白酶水解金枪鱼蒸煮液蛋白，研究不同蛋白酶对蒸煮液蛋白酶解率、分子量及氨基酸组成的影响，评价不同酶解物的促成骨细胞增殖活性并优化酶解工艺。结果表明，胰蛋白酶表现出较高的金枪鱼蒸煮液蛋白酶解率（53.51%）。酶解物的小分子肽占比达85.85%，且富含甘氨酸、酪氨酸、苯丙氨酸等促成骨细胞增殖活性特征氨基酸。胰蛋白酶酶解物的促成骨细胞增殖活性最佳，剂量-效应关系符合y=0.4139x~3-20.558x~2+49.581x+99.203(R~2=0.9597)。经正交试验优化获得胰蛋白酶最佳酶解工艺为：酶解时间3.5 h、酶解温度50℃、酶比活10000 U/g、pH8.0。金枪鱼蒸煮液蛋白在最优工艺下酶解物（1 mg/mL蛋白浓度）促成骨细胞增殖率达136.71%。因此，胰蛋白酶在生产金枪鱼成骨细胞增殖活性肽方面具有潜力。

关键词(KeyWords)： 金枪鱼蒸煮液;促成骨细胞增殖肽;水解物;工艺优化

Abstract:

Keywords:

基金项目(Foundation): 福建省农科院优秀科技创新人才专项（YCZX202410）;; 福建省属公益类科研院所基本科研专项（2022R1032004）

作者(Author): 吴羽茜,王瑛,林晓姿,陈秉彦,何志刚

DOI: 10.13386/j.issn1002-0306.2024060372

参考文献(References)：

• [1]王欣欣.金枪鱼蒸煮液主要组分的提取及生物活性研究[D].青岛：青岛科技大学,2023.[WANG X X.Extraction and bioactivity of the main components of tuna cooking liquid[D].Qingdao:Qingdao University of Science and Technology,2023.]
• [2]孙丽,夏文水.蒸煮对金枪鱼肉及其蛋白质热变性的影响[J].食品与机械，2010,26(1):22-25.[SUN L,XIA W S.Effect of steaming on thermal denaturation of tuna meat and its proteins[J].Food and Machinery,2010,26(1):22-25.]
• [3]叶孟亮.牦牛骨胶原蛋白肽抗骨质疏松作用机制研究[D].北京：中国农业科学院,2019.[YE M L.Research on the mechanism of anti-osteoporosis effect of yak collagen peptide[D].Beijing:Chinese Academy of Agricultural Sciences,2019.]
• [4]缪函霖,蒋璐,王锡昌,等.不同新鲜度金枪鱼肉蒸煮品质的研究[J].食品工业科技，2014,35(13):80-84,295.[MU H L,JIANG L,WANG X C,et al.Study on cooking quality of tuna meat with different freshness[J].Food Industry Science and Technology,2014,35(13):80-84,295.]
• [5]XU Y J,HAN X L,LI Y.Effect of marine collagen peptides on longbone development in growing rats[J].J Sci Fo od Agric,2010,90(9):1485-1491.
• [6]AKBARIAN M,KHANI A,EGHBALPOUR S,et al.Bioactive peptides:Synthesis,sources,applications,and proposed mechanisms of action[J].Int J Mol Sci,2022,23(3):14-45.
• [7]李勇.肽临床营养学[M].北京：北京大学医学出版社,2012:1-356.[LI Y.Peptide clinical nutrition[M].Beijing:Peking University Medical Press,2012.]
• [8]王珊珊.鳕鱼骨胶原肽与活性钙的制备及其抗骨质疏松活性研究[D].青岛：中国海洋大学,2013.[WANG S S.Preparation of cod collagen peptide with active calcium and its anti-osteoporosis activity[D].Qingdao:Ocean University of China,2013.]
• [9]杨溢,岳午阳,张大超,等.海洋鱼骨胶原低聚肽与天门冬氨酸钙联用增加去卵巢大鼠骨密度[J].卫生研究，2019,48(4):606-610.[YANG Y,YUE W Y,ZHANG D C,et al.Marine fish collagen oligopeptide in combination with calcium aspartate increases bone mineral density in ovariectomized rats[J].Health Research,2019,48(4):606-610.]
• [10]舒聪涵.金枪鱼骨胶原肽及其钙螯合物对成骨细胞的活性影响研究[D].舟山：浙江海洋大学,2021.[SHU C H.Effects of tuna collagen peptide and its calcium chelate on the activity of osteoblasts[D].Zhoushan:Zhejiang Ocean University,2021.]
• [11]MA W J,LIANG Z C,HE B,et al.Changes in the characteristic volatile aromatic compounds in tuna cooking liquid during fermentation and deodorization by Lactobacillus plantarum RP26 and Cyberlindnera fabianii JGM9-1[J].Food Chemistry:X,2023,20:100900.
• [12]上海市酿造科学研究所.SB/T 10317-1999蛋白酶活力测定法[S].北京：中国标准出版社,1999.[Shanghai Brewing Science Research Institute.SB/T 10317-1999 Protease activity assay[S].Beijing:China Standard Press,1999.]
• [13]中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB/T 5009.5-2016食品安全国家标准食品中蛋白质的测定[S].北京：中国标准出版社,2016.[National Health and Family Planning Commission of the People's Republic of China,China Food and Drug Administration.GB/T 5009.5-2016 National standard for food safety-Determination of protein in food[S].Beijing:China Standard Press,2016.]
• [14]全国特殊膳食标准化技术委员会.GB/T 22729-2008海洋鱼低聚肽粉[S].北京：中国标准出版社,2008.[National Technical Committee for Special Dietary Standardization.GB/T 22729-2008Marine fish oligopeptide powder[S].Beijing:China Standard Press,2008.]
• [15]贾鹏禹,孙蕊,寇芳,等.柱前衍生化液相色谱法测定牛血清中游离氨基酸含量[J].中国生物制品学杂志，2018,31(8):869-873.[JIA P Y,SUN R,KOU F,et al.Determination of free amino acids in bovine serum by pre-column derivatization liquid chromatography[J].Chinese Journal of Biological Products,2018,31(8):869-873.]
• [16]中华人民共和国国家卫生健康委员会,国家市场监督管理总局.GB 31645-2018食品安全国家标准胶原蛋白肽法[S].北京：中国标准出版社,2018.[National Health and Family Planning Commission of the People's Republic of China,China Food and Drug Administration.GB 31645-2018 Food safety national standard-Collagen peptide method[S].Beijing:China Standard Press,2018.]
• [17]UDENIGWE C C,ALUKO R E.Chemometric analysis of the amino acid requirements of antioxidant food protein hydrolysates[J].International Journal of Molecular Sciences,2011,12(5):3148-3161.
• [18]YAN L J,SUN L C,CAO K Y,et al.Type I collagen from sea cucumber (Stichopus japonicus) and the role of matrix metalloproteinase-2 in autolysis[J].Food Bioscience,2021,41:100959.
• [19]AHMED R,HAQ M,CHUN B S.Characterization of marine derived collagen extracted from the by-products of bigeye tuna(Thunnus obesus)[J].International Journal of Biological Macromolecules,2019,135:668-676.
• [20]赵珊珊,朱志伟,曾庆孝,等.不同蛋白酶酶解罗非鱼肉制备蛋白水解液的过程变化规律研究[J].现代食品科技，2008(2):115-119.[ZHAO S S,ZHU Z W,ZENG Q X,et al.Study on the change pattern of protein hydrolysate prepared from tilapia meat by enzymatic hydrolysis with different proteases[J].Modern Food Science and Technology,2008(2):115-119.]
• [21]阮晓慧,韩军岐,张润光,等.食源性生物活性肽制备工艺、功能特性及应用研究进展[J].食品与发酵工业，2016,42(6):248-253.[RUAN X H,HAN J Q,ZHANG R G,et al.Progress of food-borne bioactive peptide preparation process,functional properties and application research[J].Food and Fermentation Industry,2016,42(6):248-253.]
• [22]KARAS M.Influence of physiological and chemical factors on the absorption of bioactive peptides[J].International Journal of Food Science&Technology,2019,54(5):1486-1496.
• [23]WOUTERS A G B,ROMBOUTS I,FIERENS E,et al.Relevance of the functional properties of enzymatic plant protein hydrolysates in food systems[J].Comprehensive Reviews in Food Science and Food Safety,2016,15(4):786-800.
• [24]叶子青.菠萝蛋白酶对腊鸭品质的影响研究[D].南京：南京农业大学,2021.[YE Z Q.Effects of bromelain on the quality of preserved ducks[D].Nanjing:Nanjing Agricultural University,2021.]
• [25]李荣乔.菠萝蛋白酶酶解海湾扇贝裙边制备抗氧化肽的研究[D].保定：河北农业大学,2014.[LI R Q.Study on the preparation of antioxidant peptides from the skirts of bay scallops by enzymatic hydrolysis with bromelain[D].Baoding:Hebei Agricultural University,2014.]
• [26]吴羽茜,陈秉彦,林晓姿,等.金枪鱼蒸煮液蛋白营养评价及其功能特征氨基酸分析[J].福建农业学报，2023,38(12):1-7.[WU Y X,CHEN B Y,LIN X Z,et al.Nutritional evaluation of tuna cooking liquid protein and amino acid analysis of its functional characteristics[J].Fujian Agricultural Journal,2023,38(12):1-7.]
• [27]OHARA H,IIDA H,ITO K,et al.Effects of pro-hyp,a collagen hydrolysate-derived peptide,on hyaluronic acid synthesis using in vitro cultured synovium cells and oral ingestion of collagen hydrolysates in a guinea pig model of osteoarthritis[J].Bioscience,Biotechnology,and Biochemistry,2010,74(10):2096-2099.
• [28]GUILLERMINET F,BEAUPIED H,FABIEN-SOULE V,et al.Hydrolyzed collagen improves bone metabolism and biomechanical parameters in ovariectomized mice:An in vitro and in vivo study[J].Bone,2010,46(3):827-834.