响应面法优化海参体壁自溶条件及其影响因素分析
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摘要
目的确定海参体壁自溶最优条件,研究不同金属离子和蛋白质添加剂对海参体壁自溶过程的影响。方法以三氯乙酸(trichloroacetic acid, TCA)-可溶性寡肽含量为指标,采用中心组合设计响应面法(central composite rotatable design, CCRD)优化海参体壁自溶条件。同时,通过外源添加不同金属离子和蛋白质添加剂,考察其对自溶过程的影响。结果海参体壁自溶的最适条件为温度46℃、pH6.0、底物浓度33%、NaCl浓度2.0%和自溶时间4 h。此条件下, TCA-可溶性寡肽含量的实测值与理论预测值相近,所建立的响应面模型显著,能够较好地预测海参体壁自溶过程。终浓度为5 mmol/L条件下, Fe~(3+)、Cu~(2+)和Zn~(2+)对海参体壁自溶过程起显著抑制作用, Mn~(2+)有明显促进作用, Fe~(2+)、Ca~(2+)、Mg~(2+)和K~+则基本无影响。卵清蛋白对海参体壁自溶过程基本无影响,浓缩乳清蛋白和牛血浆蛋白对海参体壁自溶过程的影响与其浓度相关。结论本研究结果可为海参体壁自溶的抑制或利用提供一定的理论依据。
Objective To determine the optimal autolysis conditions of sea cucumber body wall and investigate the effects of different metal ions and protein additives on the autolysis. Methods The autolysis conditions of sea cucumber body wall were optimized by central composite rotatable design(CCRD) method using trichloroacetic acid(TCA)-soluble oligopeptides as the index. Meanwhile, the effects of different metal ions and protein additives on the autolysis process were investigated through exogenous addition. Results The optimal autolysis conditions were as follows: temperature was 46 ℃, pH 6.0, substrate concentration was 33%, NaCl concentration was 2.0% and autolysis time was 4 h. Under these conditions, the actual value of TCA-soluble oligopeptides was close to the predicted value. The established model was significant, and could well predict the autolysis of sea cucumber body wall. At the final concentration of 5 mmol/L, Fe~(3+), Cu~(2+) and Zn~(2+) significantly inhibited the autolysis process of the body wall of sea cucumber, Mn~(2+) significantly promoted the process, while Fe~(2+), Ca~(2+), Mg~(2+) and K~+ had no obvious effect on the autolysis process. Egg white powder had no effect on autolysis of the body wall of sea cucumber. The effect of whey protein concentrate and bovine plasma protein on autolysis process in the body wall of sea cucumber was related to its concentration. Conclusion The results of this study can provide certain theoretical basis for the inhibition or utilization of autolysis in the body wall of sea cucumber.
Objective To determine the optimal autolysis conditions of sea cucumber body wall and investigate the effects of different metal ions and protein additives on the autolysis. Methods The autolysis conditions of sea cucumber body wall were optimized by central composite rotatable design(CCRD) method using trichloroacetic acid(TCA)-soluble oligopeptides as the index. Meanwhile, the effects of different metal ions and protein additives on the autolysis process were investigated through exogenous addition. Results The optimal autolysis conditions were as follows: temperature was 46 ℃, pH 6.0, substrate concentration was 33%, NaCl concentration was 2.0% and autolysis time was 4 h. Under these conditions, the actual value of TCA-soluble oligopeptides was close to the predicted value. The established model was significant, and could well predict the autolysis of sea cucumber body wall. At the final concentration of 5 mmol/L, Fe~(3+), Cu~(2+) and Zn~(2+) significantly inhibited the autolysis process of the body wall of sea cucumber, Mn~(2+) significantly promoted the process, while Fe~(2+), Ca~(2+), Mg~(2+) and K~+ had no obvious effect on the autolysis process. Egg white powder had no effect on autolysis of the body wall of sea cucumber. The effect of whey protein concentrate and bovine plasma protein on autolysis process in the body wall of sea cucumber was related to its concentration. Conclusion The results of this study can provide certain theoretical basis for the inhibition or utilization of autolysis in the body wall of sea cucumber.
引文
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[9]季晓彤,王玱,薛鹏,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.Ji XT,Wang L,Xue P,et al.Characteristics of cathepsin K of sea cucumber stichopus japonicus and its effects on autolysis[J].J China Agric Univ,2017,22(9):75-80.
[10]王玱,年益莹,薛鹏,等.刺参2种天冬氨酸蛋白酶的酶学性质及其对自溶的影响[J].食品科学,2018,39(14):99-105.Wang L,Nian YY,Xue P,et al.Two aspartic proteases in sea cucumber(Stichopus japonicus):enzymatic properties and effect on autolysis[J].Food Sci,2018,39(14):99-105.
[11]Zhu BW,Yu JW,Zhang ZS,et al.Purification and partial characterization of an acid phosphatase from the body wall of sea cucumber Stichopus japonicus[J].Process Biochem,2009,44(8):875-879.
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[13]杜英,朱蓓薇,吴海涛,等.海参乙酰胆碱酯酶的酶学特性研究[J].食品与机械,2011,27(2):8-12.Du Y,Zhu BW,Wu HT,et al.Characteristics of acetylcholinesterase from sea cucumber[J].Food Mach,2011,27(2):8-12.
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[15]Oujifard A,Benjakul S,Ahmad M,et al.Effect of bambara groundnut protein isolate on autolysis and gel properties of surimi from threadfin bream(Nemipterus bleekeri)[J].LWT-Food Sci Technol,2012,47(2):261-266.
[16]Rawdkuen S,Benjakul S.Whey protein concentrate:Autolysis inhibition and effects on the gel properties of surimi prepared from tropical fish[J].Food Chem,2008,106(3):1077-1084.
[17]Eakpetch P,Benjakul S,Visessanguan W,et al.Autolysis of pacific white shrimp(Litopenaeus vannamei)meat:characterization and the effects of protein additives[J].J Food Sci,2008,73(2):S95-S103.
[18]孙一玮,李雪,刘春娥,等.响应面法优化双酶复合水解鳕鱼加工副产物的加工工艺[J].食品安全质量检测学报,2017,8(7):2768-2773.Sun YW,Li X,Liu CE,et al.Optimization of processing technology of double enzymatic hydrolysis of cod processing by-products by response surface methodology[J].J Food Saf Qual,2017,8(7):2768-2773.
[19]Wang C,Xu LZ,Wang X,et al.A multi-scale segmentation method of oil spills in SAR images based on JSEG and spectral clustering[J].Int JSignal Proc,2014,7(1):425-432.
[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.Zheng J,Wu HT,Dong XP,et al.Establishment of evaluation indicators for autolysis of sea cucumber based on alteration in main chemical compositions[J].J Dalian Polytechnic Univ,2010,29(6):391-395.
[21]Lowry OH,Rosenbrough NJ,Fair AL,et al.Protein measurement with the folin-phenol reagents[J].J Biol Chem,1951,193(1):265-275.
[22]启航,董秀萍,高扬,等.海参体壁粗蛋白酶的提取及酶学性质研究[J].食品研究与开发,2007,28(3):4-8.Qi H,Dong XP,Gao Y,et al.Studies on the extraction and enzymology character of the crude protease from the body wall of holothurian[J].Food Res Dev,2007,28(3):4-8.
[23]Benjakul S,Visessanguan W,Tueksuban J,et al.Effect of some protein additives on proteolysis and gel-forming ability of lizardfish(Saurida tumbil)[J].Food Hydrocoll,2004,18(3):395-401.
[24]Joglekar A,May A.Product excellence through design of experiments[J].Cereal Food World,1987,32(12):857-868.
[25]侯学敏,李林霞,张直峰,等.响应面法优化薄荷叶总黄酮提取工艺及抗氧化活性[J].食品科学,2013,34(6):124-128.Hou XM,Li LX,Zhang ZF,et al.Total flavonoids from Mentha haplocalyx Briq.Leaves:optimization of extraction process by response surface methodology and antioxidant activity[J].Food Sci,2013,34(6):124-128.
[26]徐连巧,曹剑峰,谢之英,等.响应面法优化小春花根黄酮提取工艺及其抗油脂氧化活性研究[J].食品安全质量检测学报,2018,9(5):985-992.Xu LQ,Cao JF,Xie ZY,et al.Optimization of extraction process of flavonoids from Sceptridium ternatum(thumb.)Lyon root using response surface methodology and evaluation of its antioxidant activity[J].J Food Saf Qual,2018,9(5):985-992.
[27]Yongsawatdigul J,Piyadhammaviboon P.Inhibition of autolytic activity of lizardfish surimi by proteinase inhibitors[J].Food Chem,2004,87(3):447-455.
[28]Eakpetch P,Benjakul S,Visessanguan W,et al.Autolysis of pacific white shrimp(Litopenaeus vannamei)meat:characterization and the effects of protein additives[J].J Food Sci,2008,73(2):S95-S103.
[29]Saroat R,Soottawat B.Whey protein concentrate autolysis inhibition and effects on the gel properties of surimi prepared from tropical fish[J].Food Chem,2008,106(3):1077-1084.
[30]Hunt A,Park JW,Handa A.Effect of various types of egg white on characteristics and gelation of fish myofibrillar proteins[J].J Food Sci,2009,74(9):683-692.
[31]Weerasinghe VC,Morrissey MT,Chung YC,et al.Whey protein concentrate as a proteinase inhibitor in pacific whiting surimi[J].J Food Sci,1996,61(2):367-371.
[2]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.Zheng J.Biochemical changes and antioxidative oligopeptides in the autolysis of sea cucumber Stichopus japonicus[D].Zhenjiang:Jiangsu University,2012.
[3]Thurmond F,Trotter J.Morphology and biomechanics of the microfibrillar network of sea cucumber dermis[J].J Exp Biol,1996,199(8):1817-1828.
[4]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.Ren TT,Dong XP,Zhu BW,et al.Optimization on collagen peptides preparation from sea cucumber by response surface analysis[J].Food Mach,2010,26(3):120-123.
[5]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.Fisheries and Fisheries Administration of the Ministry of Agriculture.China fishery statistical yearbook 2017[M].Beijing:China Agriculture Press,2017.
[6]Zhu BW,Zheng J,Zhang ZS,et al.Autophagy plays a potential role in the process of sea cucumber body wall“melting”induced by UV irradiation[J].Wuhan Univ J Nat Sci,2008,13(2):232-238.
[7]Sun LM,Zhu BW,Wu HT,et al.Purification and characterization of cathepsin B from the gut of the sea cucumber(Stichopus japonicus)[J].Food Sci Biotechnol,2011,20(4):919-925.
[8]Zhu BW,Zhao LL,Sun LM,et al.Purification and characterization of a cathepsin L-Like enzyme from the body wall of the sea cucumber Stichopus japonicus[J].Biosci Biotech Bioch,2008,72(6):1430-1437.
[9]季晓彤,王玱,薛鹏,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.Ji XT,Wang L,Xue P,et al.Characteristics of cathepsin K of sea cucumber stichopus japonicus and its effects on autolysis[J].J China Agric Univ,2017,22(9):75-80.
[10]王玱,年益莹,薛鹏,等.刺参2种天冬氨酸蛋白酶的酶学性质及其对自溶的影响[J].食品科学,2018,39(14):99-105.Wang L,Nian YY,Xue P,et al.Two aspartic proteases in sea cucumber(Stichopus japonicus):enzymatic properties and effect on autolysis[J].Food Sci,2018,39(14):99-105.
[11]Zhu BW,Yu JW,Zhang ZS,et al.Purification and partial characterization of an acid phosphatase from the body wall of sea cucumber Stichopus japonicus[J].Process Biochem,2009,44(8):875-879.
[12]Zhu BW,Zhao JG,Yang JF,et al.Purification and partial characterization of a novelβ-1,3-glucanase from the gut of sea cucumber Stichopus japonicus[J].Process Biochem,2008,43(10):1102-1106.
[13]杜英,朱蓓薇,吴海涛,等.海参乙酰胆碱酯酶的酶学特性研究[J].食品与机械,2011,27(2):8-12.Du Y,Zhu BW,Wu HT,et al.Characteristics of acetylcholinesterase from sea cucumber[J].Food Mach,2011,27(2):8-12.
[14]郑杰,宋志进,于笛,等.水产动物自溶研究进展[J].食品工业科技,2015,36(13):397-400.Zheng J,Song ZY,Yu D,et al.Research progress in the autolysis of aquatic animals[J].Sci Technol Food Ind,2015,36(13):397-400.
[15]Oujifard A,Benjakul S,Ahmad M,et al.Effect of bambara groundnut protein isolate on autolysis and gel properties of surimi from threadfin bream(Nemipterus bleekeri)[J].LWT-Food Sci Technol,2012,47(2):261-266.
[16]Rawdkuen S,Benjakul S.Whey protein concentrate:Autolysis inhibition and effects on the gel properties of surimi prepared from tropical fish[J].Food Chem,2008,106(3):1077-1084.
[17]Eakpetch P,Benjakul S,Visessanguan W,et al.Autolysis of pacific white shrimp(Litopenaeus vannamei)meat:characterization and the effects of protein additives[J].J Food Sci,2008,73(2):S95-S103.
[18]孙一玮,李雪,刘春娥,等.响应面法优化双酶复合水解鳕鱼加工副产物的加工工艺[J].食品安全质量检测学报,2017,8(7):2768-2773.Sun YW,Li X,Liu CE,et al.Optimization of processing technology of double enzymatic hydrolysis of cod processing by-products by response surface methodology[J].J Food Saf Qual,2017,8(7):2768-2773.
[19]Wang C,Xu LZ,Wang X,et al.A multi-scale segmentation method of oil spills in SAR images based on JSEG and spectral clustering[J].Int JSignal Proc,2014,7(1):425-432.
[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.Zheng J,Wu HT,Dong XP,et al.Establishment of evaluation indicators for autolysis of sea cucumber based on alteration in main chemical compositions[J].J Dalian Polytechnic Univ,2010,29(6):391-395.
[21]Lowry OH,Rosenbrough NJ,Fair AL,et al.Protein measurement with the folin-phenol reagents[J].J Biol Chem,1951,193(1):265-275.
[22]启航,董秀萍,高扬,等.海参体壁粗蛋白酶的提取及酶学性质研究[J].食品研究与开发,2007,28(3):4-8.Qi H,Dong XP,Gao Y,et al.Studies on the extraction and enzymology character of the crude protease from the body wall of holothurian[J].Food Res Dev,2007,28(3):4-8.
[23]Benjakul S,Visessanguan W,Tueksuban J,et al.Effect of some protein additives on proteolysis and gel-forming ability of lizardfish(Saurida tumbil)[J].Food Hydrocoll,2004,18(3):395-401.
[24]Joglekar A,May A.Product excellence through design of experiments[J].Cereal Food World,1987,32(12):857-868.
[25]侯学敏,李林霞,张直峰,等.响应面法优化薄荷叶总黄酮提取工艺及抗氧化活性[J].食品科学,2013,34(6):124-128.Hou XM,Li LX,Zhang ZF,et al.Total flavonoids from Mentha haplocalyx Briq.Leaves:optimization of extraction process by response surface methodology and antioxidant activity[J].Food Sci,2013,34(6):124-128.
[26]徐连巧,曹剑峰,谢之英,等.响应面法优化小春花根黄酮提取工艺及其抗油脂氧化活性研究[J].食品安全质量检测学报,2018,9(5):985-992.Xu LQ,Cao JF,Xie ZY,et al.Optimization of extraction process of flavonoids from Sceptridium ternatum(thumb.)Lyon root using response surface methodology and evaluation of its antioxidant activity[J].J Food Saf Qual,2018,9(5):985-992.
[27]Yongsawatdigul J,Piyadhammaviboon P.Inhibition of autolytic activity of lizardfish surimi by proteinase inhibitors[J].Food Chem,2004,87(3):447-455.
[28]Eakpetch P,Benjakul S,Visessanguan W,et al.Autolysis of pacific white shrimp(Litopenaeus vannamei)meat:characterization and the effects of protein additives[J].J Food Sci,2008,73(2):S95-S103.
[29]Saroat R,Soottawat B.Whey protein concentrate autolysis inhibition and effects on the gel properties of surimi prepared from tropical fish[J].Food Chem,2008,106(3):1077-1084.
[30]Hunt A,Park JW,Handa A.Effect of various types of egg white on characteristics and gelation of fish myofibrillar proteins[J].J Food Sci,2009,74(9):683-692.
[31]Weerasinghe VC,Morrissey MT,Chung YC,et al.Whey protein concentrate as a proteinase inhibitor in pacific whiting surimi[J].J Food Sci,1996,61(2):367-371.