鹅骨胶原蛋白钙螯合肽的分离纯化及结构鉴定  

Isolation,Purification and Structural Identification of Calcium-Chelating Peptides from Goose Bone Collagen Hydrolysate

作　　者：周名洋 何雨欣 孙杨赢[1] 潘道东[1] 曹锦轩[1] ZHOU Mingyang;HE Yuxin;SUN Yangying;PAN Daodong;CAO Jinxuan(Key Laboratory of Animal Protein Food Deep Processing Technology of Zhejiang Province,College of Food and Pharmaceutical Sciences,Ningbo University,Ningbo 315800,China)

机构地区：[1]宁波大学食品与药学学院,浙江省动物蛋白食品精深加工技术重点实验室,浙江宁波315800

出　　处：《食品科学》2020年第22期8-14,共7页Food Science

基　　金：国家水禽产业技术体系岗位科学家基金项目(CARS-42-25);宁波市科技富民项目(2017C10038)。

摘　　要：利用碱性蛋白酶酶解鹅骨胶原蛋白制备钙螯合肽,通过SephadexG-25凝胶色谱及反向高效液相色谱对钙螯合肽进行分离纯化,并采用氨基酸自动分析仪、液相色谱-串联质谱、紫外吸收光谱(ultro violet spectroscopy,UV)和傅里叶红外光谱(Fourier transform infrared spectroscopy,FTIR)进行氨基酸组成和结构鉴定。经鉴定鹅骨胶原蛋白钙螯合肽富含Glu (12.67%)和Asp (7.47%),氨基酸序列为DSYVGDEAQSKR和KLLDEGR,分子质量分别为1 353.62 u和829.47 u。经合成验证,DSYVGDEAQSKR和KLLDEGR的钙螯合能力分别为(70.08±2.20) mg/g和(68.18±1.31) mg/g,UV和FTIR结果表明DSYVGDEAQSKR和KLLDEGR与钙离子螯合后,紫外吸收强度降低、酰胺胺Ⅰ带和酰胺Ⅱ带红移和N-H键蓝移,推测多肽氨基酸残基上的羧基和氨基是螯合钙离子的主要位点。In this paper,alkaline protease was used to hydrolyze goose bone collagen to prepare calcium-chelating peptides,which were separated and purified by Sephadex G-25 column chromatography and reversed-phase high-performance liquid chromatography.The amino acid composition and structure of the purified peptides were identified by automatic amino acid analyzer,liquid chromatography-tandem mass spectrometry,ultraviolet(UV)spectroscopy and Fourier transform infrared spectroscopy.The calcium-chelating peptides were rich in Glu(12.67%)and Asp(7.47%)and were identified as DSYVGDEAQSKR and KLLDEGR,with molecular masses of 1353.62 and 829.47 u,respectively.It was proved that the calcium chelating capacities of the synthesized peptides were(70.08±2.20)and(68.18±1.31)mg/g,respectively.The intensity of UV absorption decreased,the amineⅠand amideⅡbands exhibited a red shift and the N-H band showed a blue shift.Accordingly,we speculated that the peptides may chelate calcium ions mainly through the carboxyl and amino groups of amino acid residues.

关 键 词：鹅骨胶原蛋白 钙螯合肽 液相色谱-串联质谱 氨基酸组成 紫外光谱 傅里叶红外光谱 

分 类 号：TS251.1[轻工技术与工程—农产品加工及贮藏工程]