鸡蛋壳膜多肽酶解工艺及抗氧化活性研究  

Enzymatic Hydrolysis and Antioxidant Activity of Polypeptide from Eggshell Membrane

作　　者：张伟云 张凤清[1] ZHANG Wei-yun;ZHANG Feng-qing(College of Chemistry and Life Science,Changchun University of Technology,Changchun 130012,Jilin,China)

机构地区：[1]长春工业大学化学与生命科学学院,吉林长春130012

出　　处：《食品研究与开发》2021年第21期47-55,共9页Food Research and Development

基　　金：吉林省科技发展计划项目(20170204026NY)。

摘　　要：试验以鸡蛋壳膜为原料,通过酶解法制备小分子肽,并对其生物活性进行研究。以水解度为评价指标,通过单因素试验确定蛋白酶种类、酶解时间、料液比以及酶添加量等因素对壳膜多肽水解度的影响,并通过响应面设计(Box-Behnken design,BBD)对酶解条件进行三因素三水平优化;通过高效凝胶过滤色谱测定酶解后壳膜多肽分子量的分布情况,并测定壳膜多肽对DPPH·、ABTS^(+)·、·OH、O_(2)^(-)·的清除能力。结果表明,最佳酶解工艺条件为碱性蛋白酶(AP-200a),酶添加量2.4%,料液比1∶9.5(g/mL),酶解时间5 h;在此条件下,壳膜多肽水解度为27.15%,水解后相对分子质量小于1000 Da的小分子肽所占比例为90.9%;且壳膜多肽对DPPH·、ABTS^(+)·的清除率分别为(93.03±0.51)%、(94.53±0.92)%,与V_(C)的效果相当;但对·OH和O_(2)^(-)·的清除率分别为(43.33±1.10)%和(53.40±0.70)%。因此,通过该法酶解得到的壳膜多肽中小分子肽占比较高,易被人体消化吸收,且具有一定抗氧化活性。Small molecular peptides were prepared via enzymatic hydrolysis of eggshell membrane,and their biological activities were studied.Using degree of hydrolysis as the evaluation index,the influence of protease type,enzymatic hydrolysis time,material-liquid ratio,and enzyme addition to the shell membrane polypeptide was determined through a single-factor experiment.Three factors and three levels were optimized by Box-Behnken design(BBD)for enzymatic hydrolysis condition.The molecular weight distribution of shell membrane polypeptide was determined via high efficiency gel filtration chromatography.The scavenging ability of shell membrane polypeptide to DPPH·,ABTS^(+)·,·OH,and O_(2)^(-)·was determined.The results showed that the optimum enzymatic hydrolysis conditions were as follows:alkaline protease(AP-200a),enzyme addition 2.4%,feed-liquid ratio 1∶9.5(g/mL),and enzymatic hydrolysis time 5 h.Under these conditions,the degree of hydrolysis of the shell membrane polypeptide was 27.15%;the proportion of small peptides with a relative molecular weight of less than 1000 Da after hydrolysis was 90.9%;and the scavenging ability of the shell membrane polypeptide on DPPH·and ABTS^(+)·was(93.03±0.51)%and(94.53±0.92)%,respectively,which were equivalent to the effect of V_(C).However,the elimination ability of·OH and O_(2)^(-)·were(43.33±1.10)%and(53.40±0.70)%,respectively.Therefore,the small molecular peptides obtained by enzymatic hydrolysis of shell membrane peptides were relatively high in number,easily digested,and absorbed by human body,and they had a certain antioxidant activity.

关 键 词：鸡蛋壳膜多肽 酶解 响应面优化 分子量 抗氧化活性 

分 类 号：TS201.2[轻工技术与工程—食品科学]