摘要
为高效利用大鲵加工副产物大鲵皮,对大鲵皮胶原抗氧化肽的制备及分离纯化进行研究。以1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率为评价指标评价其抗氧化活性,研究大鲵皮胶原抗氧化肽的制备工艺,确定最适工艺条件,即酶添加量8000 U/g、酶解pH 9.0、酶解温度55℃、酶解时间4 h。在此工艺条件下,大鲵皮胶原抗氧化肽DPPH自由基清除率为40.2%。采用超滤分离得到分子质量不同的3个组分,即P1(>6 kDa)、P2(3~6 kDa)、P3(<3 kDa),其中P3具有较强的抗氧化活性,DPPH自由基清除率为56.7%。在此基础上,进一步采用凝胶色谱技术分离,通过Sephadex G-25分离纯化P3,得到不同分子质量的3个组分(SP1、SP2、SP3),其中SP3具有较强的抗氧化活性,DPPH自由基清除率为72.3%。最后采用氨基酸自动分析仪技术进一步分析SP3的氨基酸组成,探寻大鲵皮胶原肽抗氧化活性与其氨基酸组成之间的内在联系。结果表明:大鲵皮胶原抗氧化肽氨基酸组成中,甘氨酸、脯氨酸含量较高,胱氨酸含量较低,这一特征符合胶原抗氧化肽的氨基酸组成特征;除甘氨酸、脯氨酸含量较高外,大鲵皮胶原蛋白抗氧化肽中还含有较高含量的具有明显抗氧化活性的丙氨酸、精氨酸、谷氨酸、亮氨酸等氨基酸,这可能是大鲵皮胶原抗氧化肽SP3具有较高抗氧化活性的原因。
In order to efficiently utilize giant salamander skin as a processing by-product,the preparation,purification and purification of antioxidant peptides from giant salamander skin collagen were studied.The 1,1-diphenyl-2-picrylhydrazyl(DPPH)free radical scavenging assay was used to evaluate the antioxidant activity.The optimal process conditions that provided maximum DPPH free radical scavenging activity(40.2%)were determined as follows:enzyme dosage 8000 U/g,pH 9.0,temperature 55℃,and hydrolysis time 4 h.Three fractions,P1(>6 kDa),P2(3–6 kDa)and P3(<3 kDa),were separated from the resultant hydrolysate by ultrafiltration.P3 had strong antioxidant activity,scavenging 56.7%of DPPH free radicals.P3 was further separated by gel permeation chromatography on Sephadex G-25 into three fractions,SP1,SP2 and SP3.SP3 had strong antioxidant activity with scavenging percentage of 72.3%against DPPH free radicals.Finally,the amino acid composition of SP3 was analyzed by an automatic amino acid analyzer to explore its relationship with the antioxidant activity of SP3.It was found that SP3 was rich in glycine and proline along with lesser amounts of cystine.In addition,SP3 contained significant amounts of amino acids with antioxidant activity such as alanine,arginine,glutamate and leucine,which may be responsible for the antioxidant activity of SP3.
作者
周艳华
郏修齐
李涛
王建文
罗庆华
ZHOU Yanhua;JIA Xiuqi;LI Tao;WANG Jianwen;LUO Qinghua(Changsha Environmental Protection Voctional College,Changsha 410004,China;Zhangjiajie Golden Salamander Biological Product Development Co.Ltd.,Zhangjiajie 427000,China;Hunan Institute of Commodity Quality Inspection,Changsha 410111,China;College of Biology and Chemical Engineering,Changsha University,Changsha 410022,China)
出处
《肉类研究》
2023年第12期1-6,共6页
Meat Research
基金
湖南省教育厅科学研究项目(21C1587)
湖南省中央引导地方科技发展资金项目(2022ZYQ044)。
关键词
中国大鲵
大鲵皮
胶原蛋白
抗氧化肽
分离纯化
Chinese giant salamander
giant salamander skin
collagen
antioxidant peptides
separation and purification
