复合酶协同超声提取藜麦皂苷及其抗氧化性
|
摘要
采用复合酶协同超声提取藜麦种皮皂苷,并对其抗氧化活性进行了测定。以藜麦皂苷的提取率为指标,考察了酶配比m(纤维素酶)∶m(果胶酶)、酶用量、酶解温度、pH、酶解时间对藜麦皂苷提取率的影响,并用响应面法进行了优化。得到最佳工艺条件为:总酶用量(以藜麦种皮质量为基准,下同)为1.5%,酶配比m(纤维素酶)∶m(果胶酶)为3∶2,酶解温度为50.5℃,pH为5.5,酶解时间为0.25 h。在该条件下,藜麦皂苷的提取率较高,达到85.32%;该法对藜麦种皮皂苷的提取率比单一纤维素酶提取率(81.56%)高4.41%,比单一果胶酶提取率(82.20%)高3.66%,比单独超声提取率(73.07%)高16.76%。采用清除1,1-二苯基-2-三硝基苯肼自由基(DPPH·)的能力,分析藜麦皂苷的抗氧化活性,结果表明,藜麦皂苷具有显著的抗氧化活性,且与皂苷的质量浓度呈剂量依赖性。
Saponins were extracted from quinoa husk using complex enzyme assisted ultrasonic technology and its antioxidant activity was measured. The effects of enzyme ratio(mass ratio of cellulase to pectinase),enzyme dosage, enzyme hydrolysis temperature, pH value and enzyme hydrolysis time on the extraction yield of quinoa saponins by single factor experiment. On the basis of the single factor experiment, response surface methodology was used to optimize the technical parameters, and the optimum parameters were as follows: total enzyme dosage(based on the mass of quinoa husk, the same below) 1.5%, enzyme ratio(mass ratio of cellulase to pectinase 3∶2), enzyme hydrolysis temperature 50.5 ℃, pH=5.5, and enzyme hydrolysis time 0.25 h. Under these optimum conditions, the extraction yield of quinoa saponins reached a maximum of 85.32%, which was 4.41% higher than that using only cellulase(81.56%), 3.66% higher than that using only pectinase(82.20%), and 16.76% higher than that using only ultrasonic extraction(73.07%).The antioxidant activity of the extracted saponins was evaluated by DPPH·. The results showed that the saponins extracted from quinoa husk had significan antioxidant activitiy, and saponins' concentration and antioxidant activity was in a dose-dependent manner.
Saponins were extracted from quinoa husk using complex enzyme assisted ultrasonic technology and its antioxidant activity was measured. The effects of enzyme ratio(mass ratio of cellulase to pectinase),enzyme dosage, enzyme hydrolysis temperature, pH value and enzyme hydrolysis time on the extraction yield of quinoa saponins by single factor experiment. On the basis of the single factor experiment, response surface methodology was used to optimize the technical parameters, and the optimum parameters were as follows: total enzyme dosage(based on the mass of quinoa husk, the same below) 1.5%, enzyme ratio(mass ratio of cellulase to pectinase 3∶2), enzyme hydrolysis temperature 50.5 ℃, pH=5.5, and enzyme hydrolysis time 0.25 h. Under these optimum conditions, the extraction yield of quinoa saponins reached a maximum of 85.32%, which was 4.41% higher than that using only cellulase(81.56%), 3.66% higher than that using only pectinase(82.20%), and 16.76% higher than that using only ultrasonic extraction(73.07%).The antioxidant activity of the extracted saponins was evaluated by DPPH·. The results showed that the saponins extracted from quinoa husk had significan antioxidant activitiy, and saponins' concentration and antioxidant activity was in a dose-dependent manner.
引文
[1]Li G,Zhu F.Molecular structure of quinoa starch[J].Carbohydrate Polymers,2017,158:124-132.
[2]Vilcacundo R,Hernández-Ledesma B.Nutritional and biological value of quinoa(Chenopodium quinoa,Willd.)[J].Current Opinion in Food Science,2017,14:1-6.
[3]Navruz-Varli S,Sanlier N.Nutritional and health benefits of quinoa(Chenopodium quinoa,Willd)[J].Journal of Cereal Science,2016,69:371-376.
[4]Li G,Zhu F.Physicochemical properties of quinoa flour as affected by starch interactions[J].Food Chemistry,2017,221:1560-1568.
[5]Lorusso A,Verni M,Montemurro M,et al.Use of fermented quinoa flour for pasta making and evaluation of the technological and nutritional features[J].LWT-Food Science and Technology,2017,78:215-221.
[6]Nickel J,Spanier L P,Botelho F T,et al.Effect of different types of processing on the total phenolic compound content,antioxidant capacity,and saponin content of Chenopodium quinoa Willd grains[J].Food Chemistry,2016,209:139-143.
[7]Nowak V,Du J,Charrondière U R.Assessment of the nutritional composition of quinoa(Chenopodium quinoa Willd.)[J].Food Chemistry,2016,193:47-54.
[8]González-Teuber M,Vilo C,Bascu?án-Godoy L.Molecular characterization of endophytic fungi associated with the roots of Chenopodium quinoa inhabiting the Atacama Desert,Chile[J].Genomics Data,2017,11:109-112.
[9]James L E A.Chapter 1 quinoa(Chenopodium quinoa,Willd.):Composition,chemistry,nutritional,and functional properties[J].Advances in Food&Nutrition Research,2009,58:1-31.
[10]Graf B L,Rojas-Silva P,Rojo L E,et al.Innovations in health value and functional food development of quinoa(Chenopodium quinoa Willd.)[J].Comprehensive Reviews in Food Science and Food Safety,2015,14(4):431-445.
[11]Kowalski R J,Medina-Meza I G,Thapa B B,et al.Extrusion processing characteristics of quinoa(Chenopodium quinoa,Willd.)var.Cherry Vanilla[J].Journal of Cereal Science,2016,70:91-98.
[12]Kuljanabhagavad T,Wink M.Biological activities and chemistry of saponins from Chenopodium quinoa,Willd[J].Phytochemistry Reviews,2009,8(2):473-490.
[13]Vermeulen K,Verspreet J,Courtin C M,et al.Reduced particle size wheat bran is butyrogenic and lowers Salmonella colonization,when added to poultry feed[J].Veterinary Microbiology,2016,198:64-71.
[14]Chen Y,Xie M Y,Gong X F.Microwave-assisted extraction used for the isolation of total triterpenoid saponins from Ganoderma atrum[J].Journal of Food Engineering,2007,81(1):162-170.
[15]Kuljanabhagavad T,Thongphasuk P,Chamulitrat W,et al.Triterpenesaponins from Chenopodium quinoa Willd[J].Phytochemistry,2008,69(9):1919-1926.
[16]Du Jingting(杜静婷),Chen Chao(陈超),Fan Sanhong(范三红).Optimization of extraction conditions for saponins from Chenopodium quinoa bran by response surface method and its antioxidant activities[J].Journal of Shanxi Agricultural Sciences(山西农业科学),2016,44(7):932-937.
[17]Wang Chenjing(王晨静),Zhao Xiwu(赵习武),Lu Guoquan(陆国权),et al.A review of characteristics and utilization of Chenopodium quinoa[J].Journal of Zhejiang A&F University(浙江农林大学学报),2014,31(2):296-301.
[18]Ghasemzadeh A,Jaafar H Z E,Rahmat A,et al.Optimization of microwave-assisted extraction of zerumbone from Zingiber zerumbet L.rhizome and evaluation of antiproliferative activity of optimized extracts[J].Chemistry Central Journal,2017,11(1).DOI:10.1186/s13065-016-0235-3.
[19]Xie X,Zhu D,Zhang W,et al.Microwave-assisted aqueous twophase extraction coupled with high performance liquid chromatography for simultaneous extraction and determination of four flavonoids in Crotalaria sessiliflora,L[J].Industrial Crops and Products,2017,95:632-642.
[20]Gao T,Zhang M,Fang Z,et al.Optimization of microwave-assisted extraction of flavonoids from young barley leaves[J].International Agrophysics,2017,31(1):45-52.
[21]Martins P F,Melo M M R D,Sarmento P,et al.Supercritical fluid extraction of sterols from Eichhornia crassipes biomass using pure and modified carbon dioxide.Enhancement of stigmasterol yield and extract concentration[J].Journal of Supercritical Fluids,2016,107:441-449.
[22]Zhang Xiujuan(张秀娟).Study on isolation,purification and antibacterial activity of antimicrobial compound from Catalpa ovata G.Don[D].Jinan:Shandong Normal University(济南:山东师范大学),2009.
[23]Rodríguez-Solana R,Salgado J M,Domínguez J M,et al.Estragole quantity optimization from fennel seeds by supercritical fluid extraction(carbon dioxide-methanol)using a Box-Behnken design.Characterization of fennel extracts[J].Industrial Crops and Products,2014,60(1):186-192.
[24]Wefers D,Gmeiner B M,Tyl C E,et al.Characterization of diferuloylated pectic polysaccharides from quinoa(Chenopodium quinoa,Willd.)[J].Phytochemistry,2015,116(1):320-328.
[25]Prasad R,Zainol M S B,Ahmad I,et al.Kinetics study of microwave assisted extraction of hypoglycemic active Compounds from Ceriops Decandra sp.leaves using ethanol:Comparison with the soxhlet extraction[J].Journal of Applied Sciences,2011,11(13):2364-2369.
[26]Fa-Tao H E,Liu G P,Zhu F T,et al.Optimization hot water extraction process of polysaccharide from Hericium erinaceus for improving the yield of polysaccharide by response surface methodology[J].Food Science and Technology,2015,1:210-215.
[27]Diao Wenchao(刁文超),Wang Ran(王然),Wang Fengwu(王凤舞),et al.Optimization of the extraction of pumpkin polysaccharides by combined ultrasonic treatment and multi-enzyme hydrolysis[J].Food Science(食品科学),2012,33(18):14-20.
[28]Liu Dayu(刘达玉),Zuo Yong(左勇).Study on extraction of dietary fiber from potato dregs by enzymatic hydrolysis[J].Science and Technology of Food Industry(食品工业科技),2005,26(5):90-92.
[29]Wei Xianya(韦献雅),Yin Liqin(殷丽琴),Zhong Chen(钟成),et al.Advances in the DPPH radical scavenging assay for antioxidant activity evaluation[J].Food Science(食品科学),2014,35(9):317-322.
[30]Madl T,Sterk H,Mittelbach M,et al.Tandem mass spectrometric analysis of a complex triterpene saponin mixture of Chenopodium quinoa[J].Journal of the American Society for Mass Spectrometry,2006,17(6):795-806.
[31]Chen Yanhui(陈燕绘),Zhou Yan(周岩),Chen Yadong(陈亚东),et al.Advances in alfalfa saponin research[J].Journal of Henan Agricultural Sciences(河南农业科学),2015,44(7):11-16.
[2]Vilcacundo R,Hernández-Ledesma B.Nutritional and biological value of quinoa(Chenopodium quinoa,Willd.)[J].Current Opinion in Food Science,2017,14:1-6.
[3]Navruz-Varli S,Sanlier N.Nutritional and health benefits of quinoa(Chenopodium quinoa,Willd)[J].Journal of Cereal Science,2016,69:371-376.
[4]Li G,Zhu F.Physicochemical properties of quinoa flour as affected by starch interactions[J].Food Chemistry,2017,221:1560-1568.
[5]Lorusso A,Verni M,Montemurro M,et al.Use of fermented quinoa flour for pasta making and evaluation of the technological and nutritional features[J].LWT-Food Science and Technology,2017,78:215-221.
[6]Nickel J,Spanier L P,Botelho F T,et al.Effect of different types of processing on the total phenolic compound content,antioxidant capacity,and saponin content of Chenopodium quinoa Willd grains[J].Food Chemistry,2016,209:139-143.
[7]Nowak V,Du J,Charrondière U R.Assessment of the nutritional composition of quinoa(Chenopodium quinoa Willd.)[J].Food Chemistry,2016,193:47-54.
[8]González-Teuber M,Vilo C,Bascu?án-Godoy L.Molecular characterization of endophytic fungi associated with the roots of Chenopodium quinoa inhabiting the Atacama Desert,Chile[J].Genomics Data,2017,11:109-112.
[9]James L E A.Chapter 1 quinoa(Chenopodium quinoa,Willd.):Composition,chemistry,nutritional,and functional properties[J].Advances in Food&Nutrition Research,2009,58:1-31.
[10]Graf B L,Rojas-Silva P,Rojo L E,et al.Innovations in health value and functional food development of quinoa(Chenopodium quinoa Willd.)[J].Comprehensive Reviews in Food Science and Food Safety,2015,14(4):431-445.
[11]Kowalski R J,Medina-Meza I G,Thapa B B,et al.Extrusion processing characteristics of quinoa(Chenopodium quinoa,Willd.)var.Cherry Vanilla[J].Journal of Cereal Science,2016,70:91-98.
[12]Kuljanabhagavad T,Wink M.Biological activities and chemistry of saponins from Chenopodium quinoa,Willd[J].Phytochemistry Reviews,2009,8(2):473-490.
[13]Vermeulen K,Verspreet J,Courtin C M,et al.Reduced particle size wheat bran is butyrogenic and lowers Salmonella colonization,when added to poultry feed[J].Veterinary Microbiology,2016,198:64-71.
[14]Chen Y,Xie M Y,Gong X F.Microwave-assisted extraction used for the isolation of total triterpenoid saponins from Ganoderma atrum[J].Journal of Food Engineering,2007,81(1):162-170.
[15]Kuljanabhagavad T,Thongphasuk P,Chamulitrat W,et al.Triterpenesaponins from Chenopodium quinoa Willd[J].Phytochemistry,2008,69(9):1919-1926.
[16]Du Jingting(杜静婷),Chen Chao(陈超),Fan Sanhong(范三红).Optimization of extraction conditions for saponins from Chenopodium quinoa bran by response surface method and its antioxidant activities[J].Journal of Shanxi Agricultural Sciences(山西农业科学),2016,44(7):932-937.
[17]Wang Chenjing(王晨静),Zhao Xiwu(赵习武),Lu Guoquan(陆国权),et al.A review of characteristics and utilization of Chenopodium quinoa[J].Journal of Zhejiang A&F University(浙江农林大学学报),2014,31(2):296-301.
[18]Ghasemzadeh A,Jaafar H Z E,Rahmat A,et al.Optimization of microwave-assisted extraction of zerumbone from Zingiber zerumbet L.rhizome and evaluation of antiproliferative activity of optimized extracts[J].Chemistry Central Journal,2017,11(1).DOI:10.1186/s13065-016-0235-3.
[19]Xie X,Zhu D,Zhang W,et al.Microwave-assisted aqueous twophase extraction coupled with high performance liquid chromatography for simultaneous extraction and determination of four flavonoids in Crotalaria sessiliflora,L[J].Industrial Crops and Products,2017,95:632-642.
[20]Gao T,Zhang M,Fang Z,et al.Optimization of microwave-assisted extraction of flavonoids from young barley leaves[J].International Agrophysics,2017,31(1):45-52.
[21]Martins P F,Melo M M R D,Sarmento P,et al.Supercritical fluid extraction of sterols from Eichhornia crassipes biomass using pure and modified carbon dioxide.Enhancement of stigmasterol yield and extract concentration[J].Journal of Supercritical Fluids,2016,107:441-449.
[22]Zhang Xiujuan(张秀娟).Study on isolation,purification and antibacterial activity of antimicrobial compound from Catalpa ovata G.Don[D].Jinan:Shandong Normal University(济南:山东师范大学),2009.
[23]Rodríguez-Solana R,Salgado J M,Domínguez J M,et al.Estragole quantity optimization from fennel seeds by supercritical fluid extraction(carbon dioxide-methanol)using a Box-Behnken design.Characterization of fennel extracts[J].Industrial Crops and Products,2014,60(1):186-192.
[24]Wefers D,Gmeiner B M,Tyl C E,et al.Characterization of diferuloylated pectic polysaccharides from quinoa(Chenopodium quinoa,Willd.)[J].Phytochemistry,2015,116(1):320-328.
[25]Prasad R,Zainol M S B,Ahmad I,et al.Kinetics study of microwave assisted extraction of hypoglycemic active Compounds from Ceriops Decandra sp.leaves using ethanol:Comparison with the soxhlet extraction[J].Journal of Applied Sciences,2011,11(13):2364-2369.
[26]Fa-Tao H E,Liu G P,Zhu F T,et al.Optimization hot water extraction process of polysaccharide from Hericium erinaceus for improving the yield of polysaccharide by response surface methodology[J].Food Science and Technology,2015,1:210-215.
[27]Diao Wenchao(刁文超),Wang Ran(王然),Wang Fengwu(王凤舞),et al.Optimization of the extraction of pumpkin polysaccharides by combined ultrasonic treatment and multi-enzyme hydrolysis[J].Food Science(食品科学),2012,33(18):14-20.
[28]Liu Dayu(刘达玉),Zuo Yong(左勇).Study on extraction of dietary fiber from potato dregs by enzymatic hydrolysis[J].Science and Technology of Food Industry(食品工业科技),2005,26(5):90-92.
[29]Wei Xianya(韦献雅),Yin Liqin(殷丽琴),Zhong Chen(钟成),et al.Advances in the DPPH radical scavenging assay for antioxidant activity evaluation[J].Food Science(食品科学),2014,35(9):317-322.
[30]Madl T,Sterk H,Mittelbach M,et al.Tandem mass spectrometric analysis of a complex triterpene saponin mixture of Chenopodium quinoa[J].Journal of the American Society for Mass Spectrometry,2006,17(6):795-806.
[31]Chen Yanhui(陈燕绘),Zhou Yan(周岩),Chen Yadong(陈亚东),et al.Advances in alfalfa saponin research[J].Journal of Henan Agricultural Sciences(河南农业科学),2015,44(7):11-16.