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

ISOLATION, PURIFICATION AND NUTRITIONAL EVALUATION OF SMALL-MOLECULAR PEPTIDE OF LONG OYSTER PROTEIN FROM ZHOUSHAN, CHINA

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。

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).