裙带菜多肽的制备及其抗氧化活性的研究
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摘要
实验以裙带菜为原料制备裙带菜蛋白,通过单因素和正交试验确定了碱性蛋白酶为裙带菜抗氧化肽制备的最佳酶,最优酶解条件为:料液比4%、酶解温度55℃、酶解时间3.5h、加酶量7500U/g、pH 10.0,该条件下制备的裙带菜多肽的还原能力最强,700nm处的吸光值为0.411。对制备的裙带菜多肽进行超滤分级,从O_2~-·清除率、·OH清除率、DPPH自由基清除率、Fe~(2+)螯合率4个方面,探究了不同分子量裙带菜多肽的抗氧化能力。结果表明:裙带菜多肽对Fe~(2+)的螯合能力和DPPH自由基的清除能力较强,对O_2~-·和·OH的清除能力相对较弱,且总体呈现低分子量肽段的抗氧化活性较强,高分子量肽段的抗氧化活性较弱的趋势。
In this study,Undaria pinnatifida protein is prepared fromUndaria pinnatifida.Alkaline protease is identified as the best enzyme for the preparation of antioxidant peptides from Undaria pinnatifida by single factor test and orthogonal test.The optimum conditions for enzymatic hydrolysis are as follows:solid-liquid ratio is 4%,enzymatic hydrolysis temperature is 55 ℃,enzymatic hydrolysis time is 3.5 h,enzyme amount is 7500 U/g and pH is 10.0.Under these conditions,the polypeptides of Undaria pinnatifida have the strongest reduction ability,and the absorbance value at 700 nm is 0.411.Ultrafiltration is used to classify the polypeptides of Undaria pinnatifida.The antioxidant ability of polypeptides from Undaria pinnatifida with different molecular weights is explored from O_2~-·scavenging rate,·OH scavenging rate,DPPH free radical scavenging rate and Fe~(2+)chelating rate.The results show that the polypeptides of Undaria pinnatifida have stronger ability to chelate Fe~(2+) and scavenge DPPH free radical,but have weaker ability to scavenge O_2~-· and ·OH.In general,the antioxidant activity of low molecular weight peptides is stronger than that of high molecular weight peptides.
In this study,Undaria pinnatifida protein is prepared fromUndaria pinnatifida.Alkaline protease is identified as the best enzyme for the preparation of antioxidant peptides from Undaria pinnatifida by single factor test and orthogonal test.The optimum conditions for enzymatic hydrolysis are as follows:solid-liquid ratio is 4%,enzymatic hydrolysis temperature is 55 ℃,enzymatic hydrolysis time is 3.5 h,enzyme amount is 7500 U/g and pH is 10.0.Under these conditions,the polypeptides of Undaria pinnatifida have the strongest reduction ability,and the absorbance value at 700 nm is 0.411.Ultrafiltration is used to classify the polypeptides of Undaria pinnatifida.The antioxidant ability of polypeptides from Undaria pinnatifida with different molecular weights is explored from O_2~-·scavenging rate,·OH scavenging rate,DPPH free radical scavenging rate and Fe~(2+)chelating rate.The results show that the polypeptides of Undaria pinnatifida have stronger ability to chelate Fe~(2+) and scavenge DPPH free radical,but have weaker ability to scavenge O_2~-· and ·OH.In general,the antioxidant activity of low molecular weight peptides is stronger than that of high molecular weight peptides.
引文
[1]周峙苗,何清,马晓宇.东海红藻龙须菜的营养成分分析及评价[J].食品科学,2010,31(9):284-287.
[2]王明艳,周琴,杨俊,等.响应面法同步优化裙带菜中蛋白和多糖提取工艺[J].食品科学,2012,33(24):66-71.
[3]Heo S J,Park P J,Park E J,et al.Antioxidant activity of enzymatic extracts from a brown seaweed Ecklonia cava by electron spin resonance spectrometry and comet assay[J].European Food Research and Technology,2005,221(1-2):41-47.
[4]于慧,李明艳,张典,等.响应面试验优化裙带菜蛋白酶解工艺及酶解液抗氧化活性[J].食品科学,2017,38(6):96-103.
[5]Sampath Kumar N S,Nazeer R A,Jaiganesh R.Purification and identification of antioxidant peptides from the skin protein hydrolysate of two marine fishes,horse mackerel(Magalaspis cordyla)and croaker(Otolithes ruber)[J].Amino Acids,2012,42(5):1641-1649.
[6]齐希光,陆晓婷,张晖,等.不同分子量黑籽瓜种子多肽抗氧化能力的研究[J].食品工业科技,2016,37(9):74-80.
[7]张友维.枯草芽孢杆菌发酵花生粕制备抗氧化肽的研究[D].无锡:江南大学,2012.
[8]郭旭.麦胚酶解产物对氧化损伤细胞的保护作用及相关肽的鉴定[D].无锡:江南大学,2013.
[9]Findlay C J,Parkin K L,Stanley D W.Differential scanning calorimetry can determine kinetics of thermal denaturation of beef muscle proteins[J].Journal of Food Biochemistry,1986,10(1):1-15.
[10]Aktas N,Kilic B.Effect of microbial transglutaminase on thermal and electrophoretic properties of ground beef[J].Food Sci Technol,2005,38(8):815-819.
[11]Udenigwe C C,Aluko R E.Chemometric analysis of the amino acid requirements of antioxidant food protein hydrolysates[J].International Journal of Molecular Sciences,2011,12(5):3148-3161.
[12]凌关庭.抗氧化食品与健康[M].北京:化学工业出版社,2004.
[13]Bondet V,Brand-Williams W,Berset C.Kinetics and mechanisms of antioxidant activity using the DPPH free radical method[J].LWT-Food Science and Technology,1997,30(6):609-615.
[14]Kaur R,Arora S,Singh B.Antioxidant activity of the phenol rich fractions of leaves of Chukrasia tabularis A.Juss[J].Bioresource Technology,2008,99(16):7692-7698.
[15]孙珍.生物质谱技术和方法在蛋白类药物序列鉴定中的应用研究[D].上海:复旦大学,2011.
[2]王明艳,周琴,杨俊,等.响应面法同步优化裙带菜中蛋白和多糖提取工艺[J].食品科学,2012,33(24):66-71.
[3]Heo S J,Park P J,Park E J,et al.Antioxidant activity of enzymatic extracts from a brown seaweed Ecklonia cava by electron spin resonance spectrometry and comet assay[J].European Food Research and Technology,2005,221(1-2):41-47.
[4]于慧,李明艳,张典,等.响应面试验优化裙带菜蛋白酶解工艺及酶解液抗氧化活性[J].食品科学,2017,38(6):96-103.
[5]Sampath Kumar N S,Nazeer R A,Jaiganesh R.Purification and identification of antioxidant peptides from the skin protein hydrolysate of two marine fishes,horse mackerel(Magalaspis cordyla)and croaker(Otolithes ruber)[J].Amino Acids,2012,42(5):1641-1649.
[6]齐希光,陆晓婷,张晖,等.不同分子量黑籽瓜种子多肽抗氧化能力的研究[J].食品工业科技,2016,37(9):74-80.
[7]张友维.枯草芽孢杆菌发酵花生粕制备抗氧化肽的研究[D].无锡:江南大学,2012.
[8]郭旭.麦胚酶解产物对氧化损伤细胞的保护作用及相关肽的鉴定[D].无锡:江南大学,2013.
[9]Findlay C J,Parkin K L,Stanley D W.Differential scanning calorimetry can determine kinetics of thermal denaturation of beef muscle proteins[J].Journal of Food Biochemistry,1986,10(1):1-15.
[10]Aktas N,Kilic B.Effect of microbial transglutaminase on thermal and electrophoretic properties of ground beef[J].Food Sci Technol,2005,38(8):815-819.
[11]Udenigwe C C,Aluko R E.Chemometric analysis of the amino acid requirements of antioxidant food protein hydrolysates[J].International Journal of Molecular Sciences,2011,12(5):3148-3161.
[12]凌关庭.抗氧化食品与健康[M].北京:化学工业出版社,2004.
[13]Bondet V,Brand-Williams W,Berset C.Kinetics and mechanisms of antioxidant activity using the DPPH free radical method[J].LWT-Food Science and Technology,1997,30(6):609-615.
[14]Kaur R,Arora S,Singh B.Antioxidant activity of the phenol rich fractions of leaves of Chukrasia tabularis A.Juss[J].Bioresource Technology,2008,99(16):7692-7698.
[15]孙珍.生物质谱技术和方法在蛋白类药物序列鉴定中的应用研究[D].上海:复旦大学,2011.