pH渐变条件下双酶分步酶解的猪皮胶原肽的体内外抗氧化活性研究  

Study on the Anti-oxidation Activity of Pigskin Collagen Peptide by Double-enzyme Step-by-step Enzymatic Hydrolysis under the Condition of pH Gradient

作　　者：张康华 姜珊 高鹏 代龙 ZHANG Kang-hua;JIANG Shan;GAO Peng;DAI Long(School of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,Shandong,China)

机构地区：[1]山东中医药大学药学院

出　　处：《食品研究与开发》2019年第20期9-13,共5页Food Research and Development

基　　金：国家科技支撑计划项目(No.2008BAI53B073);山东省自然科学基金项目(ZR2011HM051);山东省重点研发计划项目(2015GSF119012)

摘　　要：研究pH渐变条件下采用碱性蛋白酶和菠萝蛋白酶分步酶解猪皮得到的不同分子量胶原蛋白肽的体内外抗氧化活性。猪皮预处理后,采用碱性蛋白酶和菠萝蛋白酶分步进行酶解,酶解液经超滤得到M<1 kDa、1 kDa<M<3 kDa、M>3 kDa 3个不同分子量肽段,以福林酚比色法测定各组分肽含量,通过测定各组分肽段对DPPH自由基、ABTS+自由基、O2-自由基及OH自由基的清除实验评价各组分肽的体外抗氧化活性,再以不同分子量肽段酶解液对小鼠进行灌胃,以试剂盒测定小鼠灌胃后血清的总超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶的活性、总抗氧化能力和微量丙二醛含量评价各组分肽的体内抗氧化活性。综合体内外抗氧化实验数据,各组分肽的抗氧化能力强弱顺序为:小于1 kDa组分>1 kDa^3 kDa组分>大于3 kDa组分。pH渐变条件下采用碱性蛋白酶和菠萝蛋白酶分步酶解的各组分猪皮胶原蛋白肽在体内外均有不同程度的抗氧化能力,且分子量越小,抗氧化活性越强,可对机体起到保护作用。To study the antioxidant activity of collagen peptides with different molecular weight obtained from pig skin hydrolyzed by alkaline protease and bromelain step by step under the condition of pH gradual change.After pretreatment of pig skin,alkaline protease and bromelain were used to hydrolyze the enzyme step by step.Three different molecular weight peptides,M<1 kDa and 1 kDa<M<3 kDa,M>3 kDa,were obtained by ultrafiltration.The content of peptides was determined by flinophenol colorimetric method,and the scavenging effects of each peptide segment on DPPH radical,ABTS+radical,O2-radical and OH radical were evaluated.The antioxidant activity of each component peptide in vitro,and then the mice were given intragastrically with different molecular weight peptidyl hydrolysates.The activities of total SOD,Glutathion Peroxidase and catalase in serum of mice after gastric perfusion were measured by kit.Total antioxidant capacity and trace malondialdehyde content were used to evaluate the antioxidant activity of each peptide in vivo.According to the experimental data of antioxidant activity in vitro and in vivo,the order of antioxidant capacity of each peptide was smaller than that of 1 kDa component>1 kDa-3 kDa component>greater than that of 3 kDa component.Under the condition of pH gradual change,the pig skin gum was hydrolyzed by alkaline protease and bromelain step by step.Proto-protein peptides have different degrees of antioxidant capacity in vitro and in vivo,and the smaller the molecular weight,the stronger the antioxidant activity,which can play a protective role in the body.

关 键 词：双酶分步酶解 碱性蛋白酶 菠萝蛋白酶 抗氧化活性 体内外 

分 类 号：TS2[轻工技术与工程—食品科学与工程]