响应面试验优化酶法制备辣木籽多肽工艺及其抑菌活性分析
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摘要
以辣木籽蛋白粉为原料,首先筛选蛋白水解效果较佳的蛋白酶,其次考察酶添加量(酶底质量百分比)、料液比、酶解温度及酶解时间对蛋白水解度和肽得率的影响,并结合响应面法优化辣木籽多肽制备工艺,最后研究该酶解产物的抑菌活性。实验结果表明,碱性蛋白酶具有较好的蛋白水解度,酶法制备辣木籽多肽的最佳工艺条件为碱性蛋白酶添加量5.50%、pH9.0、料液比1:30(g/mL)、酶解温度62.50℃、酶解时间143 min,在此条件下肽得率实际值为(84.43±2.31)%、蛋白水解度为(20.69±0.46)%。该辣木籽蛋白酶解产物对单增李斯特菌、大肠杆菌、金黄色葡萄球菌、鼠伤寒沙门氏菌等4种致病菌株均有抑制作用,最小抑菌浓度依次为3.00 mg/mL、0.75 mg/mL、0.38 mg/mL、0.75 mg/mL,对培养至4.5 h后的金黄色葡萄球菌呈现较好抑制效果。
Moringa oleifera seed protein powder was used as raw material to prepare for peptides by enzymatic hydrolysis.Firstly,the protease with better proteolysis effect was screened as the optimal enzyme.Secondly,the effects of enzyme addition amount(enzyme-substrate mass percentage),material-liquid ratio,hydrolysis temperature and hydrolysis time on the degree of hydrolysis(DH) and peptide yield were investigated.The process for enzymatic preparation of Moringa oleifera seed peptides was optimized by response surface methodology(RSM).Finally,the antibacterial activity of the hydrolysates was evaluated.Results showed that alkaline protease was the optimal enzyme.The optimal process for enzymatic preparation of Moringa oleifera seed peptides was founded to be alkaline protease addition of 5.50%,p H of 9.0,solid-liquid ratio of 1:30(g/mL),hydrolysis temperature of 62.50 and hydrolysis time of 143 min.Under these conditions,the actual peptide ℃yield was(84.43±2.31)%,and the degree of hydrolysis was(20.69±0.46)%.The hydrolysates had inhibitory effects on Listeria monocytogenes,Escherichia coli,Staphylococcus aureus and Salmonella typhimurium,and the minimum inhibitory concentrations against these above four pathogens were 3.00 mg/mL,0.75 mg/mL,0.38 mg/mL and 0.75 mg/mL,respectively.The hydrolysates had better inhibitory effects on Staphylococcus aureus after incubation for 4.5 h.
Moringa oleifera seed protein powder was used as raw material to prepare for peptides by enzymatic hydrolysis.Firstly,the protease with better proteolysis effect was screened as the optimal enzyme.Secondly,the effects of enzyme addition amount(enzyme-substrate mass percentage),material-liquid ratio,hydrolysis temperature and hydrolysis time on the degree of hydrolysis(DH) and peptide yield were investigated.The process for enzymatic preparation of Moringa oleifera seed peptides was optimized by response surface methodology(RSM).Finally,the antibacterial activity of the hydrolysates was evaluated.Results showed that alkaline protease was the optimal enzyme.The optimal process for enzymatic preparation of Moringa oleifera seed peptides was founded to be alkaline protease addition of 5.50%,p H of 9.0,solid-liquid ratio of 1:30(g/mL),hydrolysis temperature of 62.50 and hydrolysis time of 143 min.Under these conditions,the actual peptide ℃yield was(84.43±2.31)%,and the degree of hydrolysis was(20.69±0.46)%.The hydrolysates had inhibitory effects on Listeria monocytogenes,Escherichia coli,Staphylococcus aureus and Salmonella typhimurium,and the minimum inhibitory concentrations against these above four pathogens were 3.00 mg/mL,0.75 mg/mL,0.38 mg/mL and 0.75 mg/mL,respectively.The hydrolysates had better inhibitory effects on Staphylococcus aureus after incubation for 4.5 h.
引文
[1]许敏,赵三军,宋晖,等.辣木的研究进展[J].食品科学,2016,37(23):291-301XU Min,ZHAO San-jun,SONG Hui,et al.Advances in knowledge of Moringa oleifera[J].Food Science,2016,37(23):291-301
[2]Lakshmipriya G,Kruthi D,Devarai S K.Moringa oleifera:Areview on nutritive importance and its medicinal application[J].Food Science and Human Wellness,2016,5:49-56
[3]刘凤霞,王苗苗,赵有为,等.辣木中功能性成分提取及产品开发的研究进展[J].食品科学,2015,36(19):282-286LIU Feng-xia,WANG Miao-miao,ZHAO You-wei,et al.Extraction of functional components from Moringa oleifera and development of Moringa oleifera-based products[J].Food Science,2015,36(19):282-286
[4]Brilhante R S N,Sales J A,Pereira V S.Research advances on the multiple uses of Moringa oleifera:A sustainable alternative for socially neglected population[J].Asian Pacific Journal of Tropical Medicine,2017,10(7):621-630
[5]樊建麟,邵金良,叶艳萍,等.辣木籽营养成分含量测定[J].中国食物与营养,2016,22(5):69-72FAN Jian-lin,SHAO Jin-liang,YE Yan-ping,et al.Determination on nutritional components in seeds of Moringa oleifera[J].Food and Nutrition in China,2016,22(5):69-72
[6]Ramesh K S,Iyyakkannu S,Young S K.Phytochemicals of Moringa oleifera:A review of their nutritional,therapeutic and industrial significance[J].Journal of Biotechnology,2016,6:203
[7]郭刚军,龙继明,黄艳丽,等.多油辣木不同部位营养成分分析及评价[J].食品工业科技,2016,37(22):354-364GUO Gang-jun,LONG Ji-ming,HUANG Yan-li,et al.Analysis and evaluation of nutritional components in different locations of Moringa oleifera[J].Science and Technology of Food Industry,2016,37(22):354-364
[8]Ferreira R S,Napoleao T H,Santons A F,et al.Coagulant and antibacterial activities of the water-soluble seed lectin from Moringa oleifera[J].Letters in Applied Microbiology,2011,53(2):186-192
[9]Abeyrathne E D,Lee H Y,Jo C,et al.Enzymatic hydrolysis of ovalbumin and the functional properties of the hydrolysates[J].Poultry Science,2014,93(10):2678-2686
[10]Sabrine K,Rafik B,Ali B,et al.Antibacterial activity of novel peptides isolated from protein hydrolysates of Ru Bis CO purified from green juice alfalfa[J].Journal of Functional Foods,2015,18:703-713
[11]Assaad S,Karima H,Remi P,et al.Antibacterial activity of new peptides from barbel protein hydrolysates and mode of action via a membrane damage mechanism against Listeria monocytogenes[J].Journal of Functional Foods,2014,11:322-329
[12]段琼芬,刘飞,罗金岳,等.辣木籽油的超临界CO2萃取及其化学成分分析[J].中国油脂,2010,35(2):76-79DUAN Qiong-fen,LIU Fei,LUO Jin-yue,et al.Extraction of Moringa oleifear seed oil by SCF-CO2 and analysis of its constituent[J].China Oils and Fats,2010,35(2):76-79
[13]丛艳君,于晓凤,陈澍.响应面法优化草鱼内脏蛋白质酶解工艺[J].食品科学,2015,36(10):43-48CONG Yan-jun,YU Xiao-feng,CHEN Shu.Optimization of enzymatic hydrolysis conditions of grass carp visceral proteins by response surface methodology[J].Food Science,2015,36(10):43-48
[14]高义霞,周向军,魏苇娟,等.豆渣蛋白肽的酶解工艺,抗氧化作用及其特性研究[J].中国粮油学报,2014,29(4):48-49GAO Yi-xia,ZHOU Xiang-jun,WEI Wei-juan,et al.Enzymolysis technology and antioxidant activities of bean dregs protein peptides and its partial properties[J].Journal of the Chinese Cereals and Oils Association,2014,29(4):48-49
[15]Naqash S Y,Nazeer R A.Optimization of enzymatic hydrolysis conditions for the production of antioxidant peptides from muscles of Nemipterus japonicus and Exocoetus volitans using response surface methodology[J].Amino Acids,2012,43(1):337-345
[16]祝婧,刘磊,张名位,等.不同分子量海鲈鱼胶原蛋白肽组分的功能特性比较[J].现代食品科技,2014,30(12):113-118ZHU Jing,LIU Lei,ZHANG Ming-wei,et al.Comparing the functional properties of sea bass collagen peptides(SBCP)with different molecular weights[J].Modern Food Science and Technology,2014,30(12):113-118
[17]刘振锋,刘东红,黄小鸣,等.酶法制备鱿鱼皮胶原蛋白肽工艺的条件优化[J].食品研究与开发,2016,37(5):72-76LIU Zhen-feng,DAI Sheng-jia,HUANG Xiao-ming,et al.Optimization of enzymatic preparation of collagen peptides from squid skin[J].Food Research and Development,2016,37(5):72-76
[18]易起达,王茜茜,王立峰,等.酶解制备菜籽蛋白肽条件的优化[J].食品科学,2013,9(30):167-169YI Qi-da,WANG Xi-xi,WANG Li-feng,et al.Optimization of enzymatic preparation of rapeseed peptides[J].Food Science,2013,9(30):167-169
[19]Tang W T,Zhang H,Wang L,et al.Antimicrobial peptide isolated from ovalbumin hydrolysate by immobilized liposome-binding extraction[J].European Food Research and Technology,2013,237(4):591-600
[20]Tang W T,Zhang H,Wang L,et al.New cationic antimicrobial peptide screened from boiled-dried anchovies by immobilized bacterial membrane liposome chromatography[J].Journal of Agricultural and Food Chemistry,2014,62(7):1564-1571
[21]Xiao J H,Zhang H,Shao D D.Thermodynamics of antimicrobial peptide JCpep8 binding to living staphylococcus aureus as pseudostationary phase in capillary electrochromatography and consequences for antimicrobial activity[J].Journal of Agricultural and Food Chemistry,2012,60:4535-4541
[22]黄群,杨万根,余佶,等.杜仲籽粕蛋白酶解制备抗氧化肽工艺优化[J].食品科学,2013,34(17):205-209HUANG Qun,YANG Wan-gen,YU Ji,et al.Process optimization for antioxidant peptide preparation from Eucommia seed meal protein by enzymatic hydrolysis[J].Food Science,2013,34(17):205-209
[23]张杨,胡磊,汪少芸,等.响应面优化酶解法制备蒲公英籽蛋白抗氧化肽工艺[J].食品工业科技,2016,37(5):258-262ZHANG Yang,HU Lei,WANG Shao-yun,et al.Optimization of enzymolysis technology for preparation of antioxidant peptides from dandelion seeds-derived proteins by response surface methodology[J].Science and Technology of Food Industry,2016,37(5):258-262
[24]吕小京,操德群,徐年军.响应面试验优化酶解法制备海洋微藻微拟球藻抗氧化肽工艺[J].食品科学,2018,39(6):183-188LYU Xiao-jing,CAO De-qun,XU Nian-jun.Optimization of enzymatic preparation of antioxidant peptides from protein hydrolysate of the marine microalgae nannochloropsis by response surface methodology[J].Food Science,2018,39(6):183-188
[25]余敏,黄晶晶,付瑞燕,等.响应曲面法优化酶解豆粕蛋白制备降糖肽的工艺[J].食品工业科技,2018,39(6):108-113YU Min,HUANG Jing-jing,FU Rui-yan,et al.Optimization of enzymatic hydrolysis preparation of hypoglycemic peptide from soybean meal protein by response surface methodology[J].Science and Technology of Food Industry,2018,39(6):108-113
[26]Kadam S U,Tiwari B K,Smyth T J,et al.Optimization of ultrasound assisted extraction of bioactive components from brown seaweed Ascophyllum nodosum using response surface methodology[J].Ultrasonics Sonochemistry,2015,23:308-316
[27]Muralidhar R V,Chirumamila R R,Marchanta R,et al.Aresponse surface approach for the comparison of lipase production by Candida cylindracea using two different carbon sources[J].Biochemical Engineering Journal,2001,9(1):17-23
[28]王宁,林小翠,王文君.响应面法优化黄金茶总黄酮提取工艺及对胰蛋白酶活性影响的研究[J].现代食品科技,2018,34(3):121-130WANG Ning,LIN Xiao-cui,WANG Wen-jun.Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.Hu.leaves by response surface methodology and its effect on trypsin activity[J].Modern Food Science and Technology,2018,34(3):121-130
[29]Xiao J H,Zhang H,Niu L Y,et al.Efficient screening of a novel antimicrobial peptide from Jatropha curcas by cell membrane affinity chromatography[J].Journal of Agricultural and Food Chemistry,2011,59:1145-1151
[30]杨鹏斌,于新,等.绿色木霉菌发酵液萃取物对金黄色葡萄球菌细胞膜的损伤作用[J].食品科学,2013,34(15):27-31YANG Peng-bin,YU Xin,YANG Jing,et al.Effect of extract from trichodermaviride fermentation broth on cell membrane injury of Staphylococcu saureus[J].Food Science,2013,34(15):27-31
[2]Lakshmipriya G,Kruthi D,Devarai S K.Moringa oleifera:Areview on nutritive importance and its medicinal application[J].Food Science and Human Wellness,2016,5:49-56
[3]刘凤霞,王苗苗,赵有为,等.辣木中功能性成分提取及产品开发的研究进展[J].食品科学,2015,36(19):282-286LIU Feng-xia,WANG Miao-miao,ZHAO You-wei,et al.Extraction of functional components from Moringa oleifera and development of Moringa oleifera-based products[J].Food Science,2015,36(19):282-286
[4]Brilhante R S N,Sales J A,Pereira V S.Research advances on the multiple uses of Moringa oleifera:A sustainable alternative for socially neglected population[J].Asian Pacific Journal of Tropical Medicine,2017,10(7):621-630
[5]樊建麟,邵金良,叶艳萍,等.辣木籽营养成分含量测定[J].中国食物与营养,2016,22(5):69-72FAN Jian-lin,SHAO Jin-liang,YE Yan-ping,et al.Determination on nutritional components in seeds of Moringa oleifera[J].Food and Nutrition in China,2016,22(5):69-72
[6]Ramesh K S,Iyyakkannu S,Young S K.Phytochemicals of Moringa oleifera:A review of their nutritional,therapeutic and industrial significance[J].Journal of Biotechnology,2016,6:203
[7]郭刚军,龙继明,黄艳丽,等.多油辣木不同部位营养成分分析及评价[J].食品工业科技,2016,37(22):354-364GUO Gang-jun,LONG Ji-ming,HUANG Yan-li,et al.Analysis and evaluation of nutritional components in different locations of Moringa oleifera[J].Science and Technology of Food Industry,2016,37(22):354-364
[8]Ferreira R S,Napoleao T H,Santons A F,et al.Coagulant and antibacterial activities of the water-soluble seed lectin from Moringa oleifera[J].Letters in Applied Microbiology,2011,53(2):186-192
[9]Abeyrathne E D,Lee H Y,Jo C,et al.Enzymatic hydrolysis of ovalbumin and the functional properties of the hydrolysates[J].Poultry Science,2014,93(10):2678-2686
[10]Sabrine K,Rafik B,Ali B,et al.Antibacterial activity of novel peptides isolated from protein hydrolysates of Ru Bis CO purified from green juice alfalfa[J].Journal of Functional Foods,2015,18:703-713
[11]Assaad S,Karima H,Remi P,et al.Antibacterial activity of new peptides from barbel protein hydrolysates and mode of action via a membrane damage mechanism against Listeria monocytogenes[J].Journal of Functional Foods,2014,11:322-329
[12]段琼芬,刘飞,罗金岳,等.辣木籽油的超临界CO2萃取及其化学成分分析[J].中国油脂,2010,35(2):76-79DUAN Qiong-fen,LIU Fei,LUO Jin-yue,et al.Extraction of Moringa oleifear seed oil by SCF-CO2 and analysis of its constituent[J].China Oils and Fats,2010,35(2):76-79
[13]丛艳君,于晓凤,陈澍.响应面法优化草鱼内脏蛋白质酶解工艺[J].食品科学,2015,36(10):43-48CONG Yan-jun,YU Xiao-feng,CHEN Shu.Optimization of enzymatic hydrolysis conditions of grass carp visceral proteins by response surface methodology[J].Food Science,2015,36(10):43-48
[14]高义霞,周向军,魏苇娟,等.豆渣蛋白肽的酶解工艺,抗氧化作用及其特性研究[J].中国粮油学报,2014,29(4):48-49GAO Yi-xia,ZHOU Xiang-jun,WEI Wei-juan,et al.Enzymolysis technology and antioxidant activities of bean dregs protein peptides and its partial properties[J].Journal of the Chinese Cereals and Oils Association,2014,29(4):48-49
[15]Naqash S Y,Nazeer R A.Optimization of enzymatic hydrolysis conditions for the production of antioxidant peptides from muscles of Nemipterus japonicus and Exocoetus volitans using response surface methodology[J].Amino Acids,2012,43(1):337-345
[16]祝婧,刘磊,张名位,等.不同分子量海鲈鱼胶原蛋白肽组分的功能特性比较[J].现代食品科技,2014,30(12):113-118ZHU Jing,LIU Lei,ZHANG Ming-wei,et al.Comparing the functional properties of sea bass collagen peptides(SBCP)with different molecular weights[J].Modern Food Science and Technology,2014,30(12):113-118
[17]刘振锋,刘东红,黄小鸣,等.酶法制备鱿鱼皮胶原蛋白肽工艺的条件优化[J].食品研究与开发,2016,37(5):72-76LIU Zhen-feng,DAI Sheng-jia,HUANG Xiao-ming,et al.Optimization of enzymatic preparation of collagen peptides from squid skin[J].Food Research and Development,2016,37(5):72-76
[18]易起达,王茜茜,王立峰,等.酶解制备菜籽蛋白肽条件的优化[J].食品科学,2013,9(30):167-169YI Qi-da,WANG Xi-xi,WANG Li-feng,et al.Optimization of enzymatic preparation of rapeseed peptides[J].Food Science,2013,9(30):167-169
[19]Tang W T,Zhang H,Wang L,et al.Antimicrobial peptide isolated from ovalbumin hydrolysate by immobilized liposome-binding extraction[J].European Food Research and Technology,2013,237(4):591-600
[20]Tang W T,Zhang H,Wang L,et al.New cationic antimicrobial peptide screened from boiled-dried anchovies by immobilized bacterial membrane liposome chromatography[J].Journal of Agricultural and Food Chemistry,2014,62(7):1564-1571
[21]Xiao J H,Zhang H,Shao D D.Thermodynamics of antimicrobial peptide JCpep8 binding to living staphylococcus aureus as pseudostationary phase in capillary electrochromatography and consequences for antimicrobial activity[J].Journal of Agricultural and Food Chemistry,2012,60:4535-4541
[22]黄群,杨万根,余佶,等.杜仲籽粕蛋白酶解制备抗氧化肽工艺优化[J].食品科学,2013,34(17):205-209HUANG Qun,YANG Wan-gen,YU Ji,et al.Process optimization for antioxidant peptide preparation from Eucommia seed meal protein by enzymatic hydrolysis[J].Food Science,2013,34(17):205-209
[23]张杨,胡磊,汪少芸,等.响应面优化酶解法制备蒲公英籽蛋白抗氧化肽工艺[J].食品工业科技,2016,37(5):258-262ZHANG Yang,HU Lei,WANG Shao-yun,et al.Optimization of enzymolysis technology for preparation of antioxidant peptides from dandelion seeds-derived proteins by response surface methodology[J].Science and Technology of Food Industry,2016,37(5):258-262
[24]吕小京,操德群,徐年军.响应面试验优化酶解法制备海洋微藻微拟球藻抗氧化肽工艺[J].食品科学,2018,39(6):183-188LYU Xiao-jing,CAO De-qun,XU Nian-jun.Optimization of enzymatic preparation of antioxidant peptides from protein hydrolysate of the marine microalgae nannochloropsis by response surface methodology[J].Food Science,2018,39(6):183-188
[25]余敏,黄晶晶,付瑞燕,等.响应曲面法优化酶解豆粕蛋白制备降糖肽的工艺[J].食品工业科技,2018,39(6):108-113YU Min,HUANG Jing-jing,FU Rui-yan,et al.Optimization of enzymatic hydrolysis preparation of hypoglycemic peptide from soybean meal protein by response surface methodology[J].Science and Technology of Food Industry,2018,39(6):108-113
[26]Kadam S U,Tiwari B K,Smyth T J,et al.Optimization of ultrasound assisted extraction of bioactive components from brown seaweed Ascophyllum nodosum using response surface methodology[J].Ultrasonics Sonochemistry,2015,23:308-316
[27]Muralidhar R V,Chirumamila R R,Marchanta R,et al.Aresponse surface approach for the comparison of lipase production by Candida cylindracea using two different carbon sources[J].Biochemical Engineering Journal,2001,9(1):17-23
[28]王宁,林小翠,王文君.响应面法优化黄金茶总黄酮提取工艺及对胰蛋白酶活性影响的研究[J].现代食品科技,2018,34(3):121-130WANG Ning,LIN Xiao-cui,WANG Wen-jun.Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.Hu.leaves by response surface methodology and its effect on trypsin activity[J].Modern Food Science and Technology,2018,34(3):121-130
[29]Xiao J H,Zhang H,Niu L Y,et al.Efficient screening of a novel antimicrobial peptide from Jatropha curcas by cell membrane affinity chromatography[J].Journal of Agricultural and Food Chemistry,2011,59:1145-1151
[30]杨鹏斌,于新,等.绿色木霉菌发酵液萃取物对金黄色葡萄球菌细胞膜的损伤作用[J].食品科学,2013,34(15):27-31YANG Peng-bin,YU Xin,YANG Jing,et al.Effect of extract from trichodermaviride fermentation broth on cell membrane injury of Staphylococcu saureus[J].Food Science,2013,34(15):27-31