金枪鱼蒸煮液酶解物的促成骨活性评价及功能肽挖掘

doi:10.7506/spkx1002-6630-20260301-003

食品科学 ›› 2026, Vol. 47 ›› Issue (18): 194-204.doi: 10.7506/spkx1002-6630-20260301-003

• 营养卫生 • 上一篇

金枪鱼蒸煮液酶解物的促成骨活性评价及功能肽挖掘

黄知几1,2，林晓姿1,2，吴羽茜1,2，丁梦真3，林丽娟1,2，刘芸1,2，何志刚1,2,*，陈秉彦1,2,*

（1.福建省农业科学院农产品加工研究所，福建福州 350003；2.福建省农产品（食品）加工重点实验室，福建福州 350003；3.芜湖职业技术大学食品与生物工程学院，安徽芜湖 241003）

发布日期:2026-09-29
基金资助:
福建省农业科学院优秀科技创新人才专项（YCZX202410）；福建省科学技术厅公益类科研院所专项（2024R1031002）；2024年度芜湖市科技项目（2024kj037）

Osteogenic Activity and Functional Peptides of Enzymatically Hydrolyzed Tuna Cooking Juice

HUANG Zhiji, LIN Xiaozi, WU Yuxi, DING Mengzhen, LIN Lijuan, LIU Yun, HE Zhigang, CHEN Bingyan

(1. Institute of Food Science and Technology, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China; 2. Fujian Key Laboratory of Agricultural Products (Food) Processing, Fuzhou 350003, China; 3. School of Food and Bioengineering, Wuhu Vocational Technical University, Wuhu 241003, China)

Published:2026-09-29

摘要/Abstract

摘要： 为明确金枪鱼蒸煮液酶解物的促骨细胞增殖潜力及作用机制，开发新型鱼源促骨活性肽奠定理论基础。采用胰蛋白酶水解金枪鱼蒸煮液获得具有促成骨细胞增殖活性的酶解物，通过超滤分离获得不同分子质量的酶解产物，评估不同酶解物的促成骨生物活性，利用转录组学解析其潜在促成骨细胞增殖机制，进一步通过纳流液相色谱-串联质谱与分子对接等手段鉴定出具有促骨细胞增殖活性的低聚肽序列。促成骨细胞活性评价表明，小分子酶解物可有效提高小鼠颅顶前骨细胞增殖率（135.21%），提升碱性磷酸酶活性并形成更多矿化沉积。转录组分析结果表明，小分子酶解物主要通过上调Wnt/β-连环蛋白和磷脂酰肌醇3-激酶/蛋白激酶B信号通路中Ccnd1、Ccnd3、Ccn4、Tcf7l2、F2r、Pik3r2、Pdpk1、Gnb2基因的转录水平促进骨细胞增殖。基于纳流液相色谱-串联质谱、分子对接及体外细胞实验验证手段共鉴定出6 条潜在的促成骨高活性肽段。其中DG-10（DGDAGA(Hyp)GPS）的促成骨细胞增殖活性最高且具有良好的消化稳定性，同时与表皮生长因子受体的结合稳定性（KD＝3.826×10－5 mol/L）高于VS-6（VSFYD(Hyp)，KD＝4.054×10－5 mol/L）；其与受体的结合位点Thr15、Gln16、Leu17/14、Asn12和Lys13是潜在的活性作用靶点。

关键词: 金枪鱼；促成骨细胞活性；低聚肽；细胞实验；分子对接

Abstract: This study employed trypsin to hydrolyze tuna cooking juice to obtain hydrolysates with osteogenic activity, with the aim of elucidating the osteogenic potential and mechanism of an enzymatic hydrolysate of tuna cooking juice and establishing a theoretical foundation for developing novel fish-derived osteogenic peptides. The hydrolysates with different molecular masses were separated via ultrafiltration and their osteogenic activities were evaluated. Transcriptomic analysis was then used to investigate the underlying mechanism of action. Furthermore, nanoflow liquid chromatography-tandem mass spectrometry (nano LC-MS/MS) and molecular docking were adopted to identify oligopeptides that promote osteoblast proliferation. Evaluation of osteogenic activity revealed that the low-molecular-mass hydrolysates effectively increased the proliferation rate (135.21%) and alkaline phosphatase activity of MC3T3-E1 cells, leading to increased mineral deposition. Transcriptomic analysis clarified that the low-molecular-mass hydrolysates primarily promoted osteoblast proliferation by upregulating the transcription levels of the Ccnd1, Ccnd3, Ccn4, Tcf7l2, F2r, Pik3r2, Pdpk1, and Gnb2 genes in the Wnt/β-catenin and phosphatidylinositol-3-kinase/protein kinase B signaling pathways. A total of six potential peptides with high osteogenic activity were identified through nano LC-MS/MS, molecular docking, and in vitro cell validation experiments. Among these peptides, DG-10 (DGDAGA(Hyp)GPS) exhibited the highest osteogenic activity and excellent digestive stability. Its binding affinity for epidermal growth factor receptor (EGFR) (KD = 3.826 × 10-5 mol/L) was higher than that of the peptide VS-6 (VSFYD (Hyp)) (KD = 4.054 × 10-5 mol/L), and the binding sites (Thr15, Gln16, Leu17/14, Asn12, and Lys13) on EGFR were predicted to be potential functional residues.

Key words: tuna; osteogenic bioactivity; oligopeptide; cell experiment; molecular docking

中图分类号:

TS254.1

黄知几，林晓姿，吴羽茜，丁梦真，林丽娟，刘芸，何志刚，陈秉彦. 金枪鱼蒸煮液酶解物的促成骨活性评价及功能肽挖掘[J]. 食品科学, 2026, 47(18): 194-204 https://doi.org/10.7506/spkx1002-6630-20260301-003

HUANG Zhiji1,2, LIN Xiaozi1,2, WU Yuxi1,2, DING Mengzhen3, LIN Lijuan1,2, LIU Yun1,2, HE Zhigang1,2,*, CHEN Bingyan1,2,*. Osteogenic Activity and Functional Peptides of Enzymatically Hydrolyzed Tuna Cooking Juice[J]. FOOD SCIENCE, 2026, 47(18): 194-204 https://doi.org/10.7506/spkx1002-6630-20260301-003

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金枪鱼蒸煮液酶解物的促成骨活性评价及功能肽挖掘

Osteogenic Activity and Functional Peptides of Enzymatically Hydrolyzed Tuna Cooking Juice

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