基于铜金属有机框架定向提取大米抗氧化肽及其活性研究
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(长沙理工大学食品与生物工程学院 长沙 410114)

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国家重点研发计划项目(2024YFF1105500);湖南省青年科技人才项目(2022RC1146);湖南省自然科学优秀青年基金项目(2022JJ20039)


Extraction of Antioxidant Peptides from Rice Protein Hydrolysate by a Copper Metal-organic Framework and Activities Study
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(Changsha University of Science and Technology, School of Food and Bioengineering, Changsha 410114)

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    目的:利用金属有机框架Cu-MOF对大米蛋白酶解物中的金属螯合抗氧化肽进行选择性提取,一步定向富集到活性抗氧化肽,探究大米抗氧化肽的富集机制并测定其抗氧化性能。方法:以Cu2+作为金属中心离子,刚性羧酸配体H3BTC为配体,溶剂热法合成Cu-MOF。对Cu-MOF吸附大米蛋白酶解物后的沉淀物进行解吸,然后对解吸下来的肽进行抗氧化活性测定。使用凝胶过滤色谱、反相高效液相色谱纯化解吸下来的金属螯合混合肽,对该肽采用液-质谱连用技术进行结构鉴定,使用pepsquencing等相关软件进行肽序预测;使用量子化学计算分析,预测抗氧化肽的抗氧化活性。结果:蛋白酶解物经Cu-MOF吸附解吸附后的组分,对DPPH、·O2-、·OH、ABTS自由基清除能力和Fe2+螯合能力均显著性提高,分别提高了8.198倍、7.156倍、19.417倍、7.060倍、1.257倍。其中对·O2-、·OH自由基清除和Fe2+螯合能力与阳性对照相当。Cu-MOF对蛋白酶解物的吸附是通过Cu-MOF中的羧基与蛋白酶解物的氨基产生氢键。鉴定出4条肽序:P1(T-P-G-V-F)、P2(P-A-C-L-Y)、P3(Y-L-L-L-S)和P4(P-A-E-T-F)。基于量子化学分析的计算,预测4条大米抗氧化肽都具有潜在的抗氧化性。结论:基于金属有机框架Cu-MOF提取蛋白酶解物中的抗氧化肽,是一种高效提取大米抗氧化肽的方法。

    Abstract:

    Objective: A metal-organic framework, Cu-MOF, was utilized for the selective extraction of metal-chelating antioxidant peptides from rice protein hydrolysates (RPHs), enabling one-step directional enrichment of active antioxidant peptides. The enrichment mechanism of rice antioxidant peptides was explored, and their antioxidant properties were evaluated. Methods: Cu-MOF was synthesized via a solvothermal method using Cu2+ as metal center and rigid carboxylic acid H3BTC as ligand. The desorbed metal-chelating peptide mixture was purified using gel filtration chromatography and reverse-phase high-performance liquid chromatography. The structure of the peptides was identified using liquid chromatography-mass spectrometry (LC-MS) and peptide sequence prediction using PepSquencing software. Quantum chemical calculations were performed to predict antioxidant potential of identified peptides. Results: The desorbed fractions demonstrated significant enhancement in free radical scavenging capacities (DPPH: 8.198-fold, ·O2-: 7.156-fold, ·OH: 19.417-fold, ABTS: 7.060-fold) and Fe2+ chelation ability (1.257-fold), with ·O2-/·OH scavenging and Fe2+ chelation capacities comparable to positive controls. Adsorption mechanism involved hydrogen bonding between carboxyl groups in Cu-MOF and amino groups in RPHs. Four peptide sequences were identified: P1 (T-P-G-V-F), P2 (P-A-C-L-Y), P3 (Y-L-L-S), and P4(P-A-E-T-F). Quantum chemical analysis predicted potential antioxidant activity for all four peptides. Conclusion: The extraction of antioxidant peptides from RPHs using the metal-organic framework Cu-MOF represents an efficient method for obtaining rice antioxidant peptides.

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陶会真,程静,高正杰,王旭峰,文李,程云辉,陈茂龙.基于铜金属有机框架定向提取大米抗氧化肽及其活性研究[J].中国食品学报,2025,25(3):249-264

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  • 收稿日期:2024-03-16
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  • 在线发布日期: 2025-05-23
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