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引用本文:何定芬,谢超,梁佳,梁瑞萍,李海波.基于UF、GFC及RP-HPLC的长牡蛎(Crassostrea gigas)蛋白抗衰老小分子活性肽(Zel'ner)的纯化及结构解析.海洋与湖沼,2020,51(1):141-147.
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基于UF、GFC及RP-HPLC的长牡蛎(Crassostrea gigas)蛋白抗衰老小分子活性肽(Zel'ner)的纯化及结构解析
何定芬1, 谢超2, 梁佳2, 梁瑞萍2, 李海波1
1.浙江国际海运职业技术学院 舟山 316021;2.浙江海洋大学食品与医药学院 舟山 316022
摘要:
牡蛎肉质鲜美,营养丰富,具有很高的食用价值。本研究以舟山长牡蛎(Crassostrea gigas)为研究对象,以DPPH自由基清除率为测试指标,通过UF、GFC及RP-HPLC等技术,对牡蛎蛋白酶解液进行分离纯化,获得牡蛎蛋白小分子肽样品,并对其氨基酸组成进行营养性评价。研究结果表明:浓度为30%-100%的硫酸铵处理酶解液的分离效果明显优于低于浓度为30%硫酸铵的分离效果,其DPPH自由基清除率达到52.43%;纯化后得到两种牡蛎蛋白小分子肽分别标记为组分a(Mw <1kDa)和组分b(1kDa < Mw < 3kDa);组分a中牡蛎蛋白小分子肽分子量分布主要为1995.2Da、1258.9Da、1023.3Da、398.1Da;对牡蛎蛋白小分子肽组分a进行检测,发现其中氨基酸种类及含量丰富,其中必需氨基酸(EAA)含量为18.863%,总氨基酸含量(TAA)为43.3748%,必需氨基酸占总氨基酸43.49%,必需氨基酸与非必需氨基酸(NEAA)的比例为76.95%,风味氨基酸(FAA)含量为18.9147%,占总氨基酸含量的43.61%。综上可见,牡蛎蛋白小分子肽具有极高的营养价值和市场经济效益,此研究为牡蛎蛋白抗衰老小分子活性肽(Zel'ner)的开发提供了新的思路和方向。
关键词:  长牡蛎(Crassostrea gigas)  小分子肽(Zel'ner)  分离纯化  氨基酸分析
DOI:10.11693/hyhz20190700143
分类号:TS254;Q956
基金项目:浙江省基础公益基金项目,LGN19C200016号;2018年国家级大学生创新创业训练计划项目,201810340044S号。
ISOLATION, PURIFICATION AND NUTRITIONAL EVALUATION OF SMALL-MOLECULAR PEPTIDE OF LONG OYSTER PROTEIN FROM ZHOUSHAN, CHINA
HE Ding-Fen1, XIE Chao2, LIANG Jia2, LIANG Rui-Ping2, LI Hai-Bo1
1.Zhejiang International Maritime Vocational and Technical College, Zhoushan 316021, China;2.College of Food and Medicine, Zhejiang Ocean University, Zhoushan 316022, China
Abstract:
Crassostrea gigas was used as the research object, and the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging rate was used as the test index. The oyster protein hydrolysate was separated and purified by UF, GFC, and RP-HPLC techniques to obtain small-molecular peptide from oyster protein (the SMP), for which nutrition of their amino acid composition was evaluated. The results show that the separation of ammonium sulfate treatment solution in concentration of 30%-100% was better than that of <30% ammonium sulfate and the DPPH free radical scavenging ratereached 52.43%. Two types of the SMP were obtained and labeled as Component a (Mw <1kDa) and Component b (1kDa < Mw < 3kDa), respectively. The molecular weight distribution of the SMP in Component a is mainly 1995.2, 1258.9, 1023.3, and 398.1Da. Component a was found rich in amino acid species and content, e.g., essential amino acid (EAA) content:18.86; total amino acid content (TAA):43.37%; essential amino acids to the total amino acids:43.49%, the ratio of essential amino acids to non-essential amino acids (NEAA):76.95%; and the content of flavor amino acids (FAA):18.91%, accounting for 43.61% of the total amino acid. Therefore, the SMP features with high nutritional and commercial value, providing us new ideas and directions for exploration and development of oyster protein anti-aging small-molecule active peptide (Zel'ner).
Key words:  Crassostrea gigas  small-molecular peptide  separation and purification  amino acid analysis
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