海参体壁自溶的响应面优化及其体外抗氧化活性研究
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摘要
采用响应面分析法优化海参体壁的自溶条件,并对其自溶水解物进行体外抗氧化活性分析。以TCA-可溶性寡肽含量为指标,利用CCRD中心组合设计研究了温度、pH和底物浓度对海参体壁自溶过程的影响。结果表明,所建立的响应面模型极显著(P <0.0001),可以较好的反映海参体壁自溶过程中各因素与TCA-可溶性寡肽含量之间的关系,且由模型得出海参体壁自溶的最适条件为温度45℃、pH 6.8、底物浓度17.70%、NaCl浓度2.0%和自溶时间4 h。在此条件下,海参体壁自溶水解物中TCA-可溶性寡肽的含量为22.98mg/g。其体外抗氧化活性分析结果表明,海参体壁自溶水解物具有一定的DPPH自由基、羟自由基和超氧阴离子自由基清除能力以及Fe2+螯合能力,其清除率和螯合率达到50%时的质量浓度分别为15.95mg/mL、0.15mg/mL、39.38mg/mL以及0.08mg/mL。抗氧化能力强弱次序依次为Fe2+螯合能力>羟自由基清除能力>DPPH自由基清除能力>超氧阴离子自由基清除能力。
The response surface methodology was used to optimize the autolysis of sea cucumber body wall,and the antioxidant activities of autolytic hydrolysate were further studied.The content of TCA-soluble oligopeptide was selected as the index,the effects of temperature,pH value and substrate concentration on the autolysis process of sea cucumber body wall were detailedly investigated by the central composite rotatable design method.The results showed that the model established was very significant(P<0.0001),and the optimal autolysis conditions were determined to be temperature 45℃,pH 6.8,substrate concentration 17.70%,NaCl concentration 2.0%and autolysis time 4h.Under these conditions,the content of TCA-soluble oligopeptide in the autolytic hydrolysate was 22.98mg/g.The autolytic hydrolysate from sea cucumber body wall possessed DPPH radical scavenging activity,OH·scavenging activity,O_(2·)~- scavenging ability and Fe~(2+)-chelating activity with IC_(50) values of 15.95,0.15,39.38 and 0.08 mg/mL,respectively.The antioxidant activities were ranked as follows:Fe~(2+)-chelating activity>OH·scavenging activity>DPPH radical scavenging activity>O_(2·)~-scavenging ability.
The response surface methodology was used to optimize the autolysis of sea cucumber body wall,and the antioxidant activities of autolytic hydrolysate were further studied.The content of TCA-soluble oligopeptide was selected as the index,the effects of temperature,pH value and substrate concentration on the autolysis process of sea cucumber body wall were detailedly investigated by the central composite rotatable design method.The results showed that the model established was very significant(P<0.0001),and the optimal autolysis conditions were determined to be temperature 45℃,pH 6.8,substrate concentration 17.70%,NaCl concentration 2.0%and autolysis time 4h.Under these conditions,the content of TCA-soluble oligopeptide in the autolytic hydrolysate was 22.98mg/g.The autolytic hydrolysate from sea cucumber body wall possessed DPPH radical scavenging activity,OH·scavenging activity,O_(2·)~- scavenging ability and Fe~(2+)-chelating activity with IC_(50) values of 15.95,0.15,39.38 and 0.08 mg/mL,respectively.The antioxidant activities were ranked as follows:Fe~(2+)-chelating activity>OH·scavenging activity>DPPH radical scavenging activity>O_(2·)~-scavenging ability.
引文
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[23]HSU B,COUPAR I M,Ng K.Antioxidant avtivity of hot water extract from the fruit of the Doum palm,Hyphaene thebaica[J].Food chemistry,2006,98(2):317-328.
[24]张磊,王锦旭,杨贤庆,等.响应面法优化合浦珠母贝糖胺聚糖咀嚼片的制备工艺[J].食品工业科技,http://kns.cnki.net/kcms/detail/11.1759.TS.20180416.1711.038.html.
[25]CHEN R Z,JIN C G,TONG Z G,et al.Optimization extraction,characterization and antioxidant activities of pectic polysaccharide from tangerine peels[J].Carbohydrate Polymers,2016,136(3):187-197.
[26]GORDON M.Antioxidant in food[M].New York-CRC Press,2001:7-21.
[27]郑捷,孙凯华,高芯,等.海参斑软骨多糖的提取及其体外抗氧化活性分析[J].中国食品添加剂,2018(7):114-121.
[28]唐仕荣,宋慧,陈尚龙,等.不同方法提取余甘子多酚的抗氧化活性及其指纹图谱分析[J].中国食品添加剂,2018(8):90-97.
[2]Sae-leaw T,O,callaghan Y C,Benjakul S,et al.Antioxidant,immunomodulatory and antiproliferative effects of gelatin hydrolysates from seabass(Lates calcarifer)skins[J].International Journal of Food Science and Technology,2016,51(7):1545-1551.
[3]刘文颖,徐姗姗,谷瑞增,等.海洋胶原低聚肽中抗氧化肽的分离及鉴定[J].食品工业科技,2017,38(6):101-105,109.
[4]ZHOU X Q,WANG C H,JIANG A L.Antioxidant peptides isolated from sea cucumber Stichopus Japonicus[J].European Food Research and Technology,2012,234(3):441-447.
[5]郑杰.海参自溶过程中生化变化及抗氧化活性寡肽的研究[D].镇江:江苏大学,2012.
[6]ASHA K K,REMYAKUMARI K R,KUMAR K A,et al.Sequence determination of an antioxidant peptide obtained by enzymatic hydrolysis of oyster crassostrea madrasensis(Preston)[J].International Journal of Peptide Research and Therapeutics,2016,22(3):421-433.
[7]JAIGANESH R,NAZEER R A,KUMAR N S S.Purification and Identification of antioxidant peptide from black pomfret,Parastromateus niger(Bloch,1975)viscera protein hydrolysate[J].Food Science and Biotechnology,2011,20(4):1087-1094.
[8]WU H T,JIN W G,SUN S G,et al.Identification of antioxidant peptides from protein hydrolysates of scallop(Patinopecten yessoensis)female gonads[J].European Food Research and Technology,2016,242(5):1-10.
[9]SUETSUNA K.Antioxidant peptides from the protease digest of prawn(Penaeus japonicus)muscle.Marine Biotechnology,2000,2(1):5-10.
[10]朱蓓薇.海珍品加工理论与技术的研究[M].北京:科学出版社,2010.
[11]农业部渔业渔政管理局.2017中国渔业统计年鉴[M].北京:中国农业出版社,2017.
[12]ZHU B W,ZHENG J,ZHANG Z S,et al.Autophagy plays a potential role in the process of sea cucumber body wall“melting”induced by UV irradiation[J].Wuhan University Journal of Natural Sciences,2008,13(2):232-238.
[13]SUN L M,ZHU B W,WU H T,et al.Purification and characterization of cathepsin B from the gut of the sea cucumber(Stichopus japonicus)[J].Food Science and Biotechnology,2011,20(4):919-925.
[14]ZHU B W,ZHAO L L,SUN L M,et al.Purification and characterization of a cathepsin L-Like enzyme from the body wall of the sea cucumber Stichopus japonicus[J].Bioscience Biotechnology and Biochemistry,2008,72(6):1430-1437.
[15]季晓彤,王玲,吴海涛,等.海参组织蛋白酶K的部分酶学性质及其对海参自溶的影响[J].中国农业大学学报,2017,22(9):75-80.
[16]ZHU B W,YU J W,ZHANG Z S,et al.Purification and partial characterization of an acid phosphatase from the body wall of sea cucumber Stichopus japonicus[J].Process Biochemistry,2009,44(8):875-879.
[17]ZHU B W,ZHAO J G,YANG J F,et al.Purification and partial characterization of a novelβ-1,3-glucanase from the gut of sea cucumber Stichopus japonicus[J].Process Biochemistry,2008,43(10):1102-1106.
[18]马冬冬.刺参自溶过程中体壁糖蛋白的变化及机理研究[D].大连:大连工业大学,2017.
[19]任婷婷,董秀萍,朱蓓薇,等.响应面法优化海参胶原蛋白肽的制备条件[J].食品与机械,2010,26(3):120-123.
[20]郑杰,吴海涛,董秀萍,等.基于主要化学成分变化建立的海参自溶评价指标[J].大连工业大学学报,2010,29(6):391-395.
[21]LOWRY OH,ROSENBROUGH N J,FAIR A L,et al.Protein measurement with the folin-phenol reagents[J].Journal of Biological Chemistry,1951,193(1):265-275.
[22]郑杰,于笛,陈冲,等.海蜇生殖腺酶解物抗氧化活性的研究[J].水产科学,2014,33(2):81-86.
[23]HSU B,COUPAR I M,Ng K.Antioxidant avtivity of hot water extract from the fruit of the Doum palm,Hyphaene thebaica[J].Food chemistry,2006,98(2):317-328.
[24]张磊,王锦旭,杨贤庆,等.响应面法优化合浦珠母贝糖胺聚糖咀嚼片的制备工艺[J].食品工业科技,http://kns.cnki.net/kcms/detail/11.1759.TS.20180416.1711.038.html.
[25]CHEN R Z,JIN C G,TONG Z G,et al.Optimization extraction,characterization and antioxidant activities of pectic polysaccharide from tangerine peels[J].Carbohydrate Polymers,2016,136(3):187-197.
[26]GORDON M.Antioxidant in food[M].New York-CRC Press,2001:7-21.
[27]郑捷,孙凯华,高芯,等.海参斑软骨多糖的提取及其体外抗氧化活性分析[J].中国食品添加剂,2018(7):114-121.
[28]唐仕荣,宋慧,陈尚龙,等.不同方法提取余甘子多酚的抗氧化活性及其指纹图谱分析[J].中国食品添加剂,2018(8):90-97.