响应面优化羊肚菌多糖提取工艺及抗氧化性
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摘要
采用超声辅助酶法提取羊肚菌多糖,探讨液料比、酶解pH、酶解时间、超声时间、酶用量对多糖得率的影响,通过Plackett-Burman试验和响应面法优化提取工艺,并研究其抗氧化活性。结果表明:液料比22∶1 mL/g、酶解pH4.0、酶解时间60 min、超声时间19 min、酶用量0.8%为最佳提取条件,此时多糖实际得率为7.79%。TCA法蛋白脱除率为77.27%、多糖保留率为72.62%。AB-8大孔树脂的脱色率为84.25%,多糖保留率为95.71%。羊肚菌多糖对DPPH自由基、羟自由基、超氧阴离子清除率最高分别为60.67%、70.77%、72.76%。综上所述,提取的多糖具有较好的抗氧化性能,本实验为羊肚菌开发成新型膳食补充剂或功能性食品提供理论依据。
To research polysaccharide by enzymes-ultrasound assisted extraction technique and determined its antioxidant activity,the liquid-solid ratio,enzymatic hydrolysis pH,enzymatic hydrolysis time,ultrasonic time and enzyme dosage were optimized by Plackett-Burman experiment and response surface methodology. The results showed that the optimized technological parameters were as follows: Liquid-solid ratio 22 ∶ 1 mL/g,enzymatic pH4.0,the enzymatic time 60 min,ultrasonic time 19 min,and the enzyme dosage 0.8%. Under these conditions,the actual crude polysaccharides yields was7.79%. The protein removal rate by TCA method and the polysaccharides retention rate were 77.27% and 72.62%,respectively.The decolorization rate of AB-8 resin was 84.25% and the retention rate of polysaccharides was 95.71%. The scavenging activities of polysaccharides from Morchella esculenta on·DPPH,·OH and O2-·were reached 60.67%,70.77% and72.76%,respectively.The extracted polysaccharides had better antioxidant properties and could be developed into a new type of dietary supplement or functional food.
To research polysaccharide by enzymes-ultrasound assisted extraction technique and determined its antioxidant activity,the liquid-solid ratio,enzymatic hydrolysis pH,enzymatic hydrolysis time,ultrasonic time and enzyme dosage were optimized by Plackett-Burman experiment and response surface methodology. The results showed that the optimized technological parameters were as follows: Liquid-solid ratio 22 ∶ 1 mL/g,enzymatic pH4.0,the enzymatic time 60 min,ultrasonic time 19 min,and the enzyme dosage 0.8%. Under these conditions,the actual crude polysaccharides yields was7.79%. The protein removal rate by TCA method and the polysaccharides retention rate were 77.27% and 72.62%,respectively.The decolorization rate of AB-8 resin was 84.25% and the retention rate of polysaccharides was 95.71%. The scavenging activities of polysaccharides from Morchella esculenta on·DPPH,·OH and O2-·were reached 60.67%,70.77% and72.76%,respectively.The extracted polysaccharides had better antioxidant properties and could be developed into a new type of dietary supplement or functional food.
引文
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[3]李慧芬,李翠妮,徐红星,等.菌类多糖应用研究进展[J].陕西农业科学,2006(5):77-80.
[4]李丽波.羊肚菌抗疲劳作用研究[J].中国野生植物资源,2001,20(1):17-18,32.
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[6]杨芳,王新风,翁良,等.两种羊肚菌胞内多糖体外抗氧化性[J].食品科学,2010,31(23):76-78.
[7]龙正海,纪其雄,周惠燕,等.羊肚菌多糖体外抗菌活性研究[J].时珍国医国药,2006,17(6):902-903.
[8]李谣,陈金龙,王丽颖,等.羊肚菌多糖抑制人乳腺癌细胞MDA-MB-231增殖和诱导细胞凋亡研究[J].食品科学,2016,37(21):214-218.
[9]贾建会,吕晓莲,樊利青,等.深层发酵羊肚菌多糖的提取分离及纯化研究[J].食品科学,2002,23(4):59-63.
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[11]王振斌,刘加友,马海乐,等.无花果多糖提取工艺优化及其超声波改性[J].农业工程学报,2014(10):262-269.
[12]肖宝石,吕海涛.酶法提取浒苔多糖工艺优化的研究[J].食品与机械,2010,26(5):125-127.
[13]Chen G J,Bu F,Chen X H,et al. Ultrasonic extraction,structural characterization, physicochemical properties and antioxidant activities of polysaccharides from bamboo shoots(Chimonobambusa quadrangularis)processing by-products[J].International Journal of Biological Macro molecules,2018,112:656-666.
[14]朱俊玲.超临界流体萃取芦荟多糖的研究[J].农产品加工·综合刊,2011(7):67-68.
[15]谢慧明,李超,潘见,等.Fe3O4-PSF磁性复合超滤膜分离麦冬多糖[J].食品科学,2010,31(22):41-45.
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[18]晁红娟,高荫榆,丁红秀,等.酶法中试提取毛竹叶、柄中多糖和类黄酮的研究[J].食品科学,2007,28(12):208-211.
[19]韩鹏远,柴美清,陈斌,等.山西中南部地区羊肚菌资源及生境调查[J].山西农业科学,2018,46(3):426-428.
[20]程振玉,杨英杰,刘治刚.超声波辅助酶法提取北五味子多糖工艺研究[J].中国酿造,2014,33(3):104-108.
[21]杜娟,时文静.香菇、金针菇、黑木耳多糖的提取与测定[J].江苏农业科学,2016,44(8):347-349,350.
[22]John W M. Nesting plackett-burman designs[J]. Statistics and Probability Letters,1996,27(3):221-223.
[23]Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry,1976,72(1):248-254.
[24]杨云,冯卫生,雷高明,等.大枣渣多糖精制纯化工艺的研究[J].中药材,2006,29(1):78-80.
[25]车向前,常明泉,陈芳,等.H2O2对白及多糖脱色工艺研究[J].实用药物与临床,2018,21(1):70-73.
[26]余钰骢.灵芝结构多糖水解物提取分离纯化和免疫功能研究[D].杭州:浙江大学,2014.
[27]Ma C W,Feng M Y,Zhai X F,et al. Optimization for the extraction of polysaccharides from Ganoderma lucidum and their antioxidant and antiproliferative activities[J]. Journal of the Taiwan Institute of Chemical Engineers,2013,44(6):886-894.
[28]梁绍兰,覃冬,黄连秋,等.柑橘皮多糖抗氧化性研究[J].安徽农业科学,2012(5):2624-2625.
[29]张燕平,张虹,洪泳平,等.羊栖菜提取物体外自由基清除能力的研究[J].郑州工程学院学报,2003,24(1):50-53,57.
[30]董玉玮,苗敬芝,曹泽虹,等.酶法提取灵芝多糖的工艺研究[J].食品科技,2009,34(6):217-220.
[31]孟雅红,李辉.Plackett-Burman设计和响应面法优化超声协同酶法提取鸡油菌多糖工艺[J].食品工业科技,2015,36(21):242-248.
[32]陈金娥,高瑞苑,张海容,等.响应面法优化胡麻籽壳多糖超声提取工艺[J].中国油脂,2017,42(9):145-148,156.
[33]尹秀莲,游庆红.超声辅助复合酶法提取桑黄多糖[J].食品与机械,2011,27(4):58-60.
[34]黎英,陈雪梅,严月萍,等.超声波辅助酶法提取红腰豆多糖工艺优化[J].农业工程学报,2015(15):293-301.
[35]陈文超,张锦华,叶峥,等.响应面法优化超声波辅助提取苹果渣低聚糖的工艺[J].食品工业科技,2017,38(12):277-281,286.
[36]洪晶,陈涛涛,唐梦茹,等.响应面法优化韭菜籽蛋白质提取工艺[J].中国食品学报,2013,13(12):89-96.
[37]Zhang X Y,Chen Z X,Wang L. Removal of protein from polysaccharides of rice bran[J]. Science&Technology of Food Industry,2008,29(3):163-165.
[38]Xu H D,Li J,Li Y J,et al.Using macroporous resin AB-8for pigment removal from onion polysaccharides[J].Food Science,2012,33(6):127-131.
[39]付丽红,王艳萍,杨延瑞,等.羊肚菌胞外多糖理化性质及抗氧化活性研究[J].食品科技,2013(11):183-188.
[40]黄俊丽,张慜.松茸、黑牛肝菌、双孢白蘑菇提取物体外抗氧化性试验研究[J].食品与生物技术学报,2011,30(3):342-347.
[41]Zhao B T,Liu J,Chen X,et al.Purification,structure and anti-oxidation of polysaccharides from the fruit of Nitraria tangutorum Bobr[J]. RSC Advances,2018,8(21):11731-11743.
[42]袁红波,张劲松,贾薇,等.利用大孔树脂对低分子量灵芝多糖脱色的研究[J].食品工业科技,2009,30(3):204-206.
[2]李娟,王臻,姚良同,等.羊肚菌多糖研究进展[J].微生物学杂志,2005,25(4):89-91.
[3]李慧芬,李翠妮,徐红星,等.菌类多糖应用研究进展[J].陕西农业科学,2006(5):77-80.
[4]李丽波.羊肚菌抗疲劳作用研究[J].中国野生植物资源,2001,20(1):17-18,32.
[5]陈彦,潘见,周丽伟,等.羊肚菌胞外多糖抗肿瘤作用的研究[J].食品科学,2008,29(9):553-556.
[6]杨芳,王新风,翁良,等.两种羊肚菌胞内多糖体外抗氧化性[J].食品科学,2010,31(23):76-78.
[7]龙正海,纪其雄,周惠燕,等.羊肚菌多糖体外抗菌活性研究[J].时珍国医国药,2006,17(6):902-903.
[8]李谣,陈金龙,王丽颖,等.羊肚菌多糖抑制人乳腺癌细胞MDA-MB-231增殖和诱导细胞凋亡研究[J].食品科学,2016,37(21):214-218.
[9]贾建会,吕晓莲,樊利青,等.深层发酵羊肚菌多糖的提取分离及纯化研究[J].食品科学,2002,23(4):59-63.
[10] Su C A,Xu X Y,Liu D Y,et al. Isolation and characterization of exopolysaccharide with immunomodulatory activity from fermentation broth of Morchella conica[J]. DARU Journal of Pharmaceutical Sciences,2013,21(1):1-6.
[11]王振斌,刘加友,马海乐,等.无花果多糖提取工艺优化及其超声波改性[J].农业工程学报,2014(10):262-269.
[12]肖宝石,吕海涛.酶法提取浒苔多糖工艺优化的研究[J].食品与机械,2010,26(5):125-127.
[13]Chen G J,Bu F,Chen X H,et al. Ultrasonic extraction,structural characterization, physicochemical properties and antioxidant activities of polysaccharides from bamboo shoots(Chimonobambusa quadrangularis)processing by-products[J].International Journal of Biological Macro molecules,2018,112:656-666.
[14]朱俊玲.超临界流体萃取芦荟多糖的研究[J].农产品加工·综合刊,2011(7):67-68.
[15]谢慧明,李超,潘见,等.Fe3O4-PSF磁性复合超滤膜分离麦冬多糖[J].食品科学,2010,31(22):41-45.
[16]明建.云南特有食药用菌活性多糖研究[D].重庆:西南大学,2008.
[17]Xu N,Sun Y H,Guo X L,et al.Optimization of ultrasonicmicrowave synergistic extraction of polysaccharides from Morchella conica[J]. Journal of Food Processing&Preservation,2018,42(2):e13423.1-e13423.7.
[18]晁红娟,高荫榆,丁红秀,等.酶法中试提取毛竹叶、柄中多糖和类黄酮的研究[J].食品科学,2007,28(12):208-211.
[19]韩鹏远,柴美清,陈斌,等.山西中南部地区羊肚菌资源及生境调查[J].山西农业科学,2018,46(3):426-428.
[20]程振玉,杨英杰,刘治刚.超声波辅助酶法提取北五味子多糖工艺研究[J].中国酿造,2014,33(3):104-108.
[21]杜娟,时文静.香菇、金针菇、黑木耳多糖的提取与测定[J].江苏农业科学,2016,44(8):347-349,350.
[22]John W M. Nesting plackett-burman designs[J]. Statistics and Probability Letters,1996,27(3):221-223.
[23]Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry,1976,72(1):248-254.
[24]杨云,冯卫生,雷高明,等.大枣渣多糖精制纯化工艺的研究[J].中药材,2006,29(1):78-80.
[25]车向前,常明泉,陈芳,等.H2O2对白及多糖脱色工艺研究[J].实用药物与临床,2018,21(1):70-73.
[26]余钰骢.灵芝结构多糖水解物提取分离纯化和免疫功能研究[D].杭州:浙江大学,2014.
[27]Ma C W,Feng M Y,Zhai X F,et al. Optimization for the extraction of polysaccharides from Ganoderma lucidum and their antioxidant and antiproliferative activities[J]. Journal of the Taiwan Institute of Chemical Engineers,2013,44(6):886-894.
[28]梁绍兰,覃冬,黄连秋,等.柑橘皮多糖抗氧化性研究[J].安徽农业科学,2012(5):2624-2625.
[29]张燕平,张虹,洪泳平,等.羊栖菜提取物体外自由基清除能力的研究[J].郑州工程学院学报,2003,24(1):50-53,57.
[30]董玉玮,苗敬芝,曹泽虹,等.酶法提取灵芝多糖的工艺研究[J].食品科技,2009,34(6):217-220.
[31]孟雅红,李辉.Plackett-Burman设计和响应面法优化超声协同酶法提取鸡油菌多糖工艺[J].食品工业科技,2015,36(21):242-248.
[32]陈金娥,高瑞苑,张海容,等.响应面法优化胡麻籽壳多糖超声提取工艺[J].中国油脂,2017,42(9):145-148,156.
[33]尹秀莲,游庆红.超声辅助复合酶法提取桑黄多糖[J].食品与机械,2011,27(4):58-60.
[34]黎英,陈雪梅,严月萍,等.超声波辅助酶法提取红腰豆多糖工艺优化[J].农业工程学报,2015(15):293-301.
[35]陈文超,张锦华,叶峥,等.响应面法优化超声波辅助提取苹果渣低聚糖的工艺[J].食品工业科技,2017,38(12):277-281,286.
[36]洪晶,陈涛涛,唐梦茹,等.响应面法优化韭菜籽蛋白质提取工艺[J].中国食品学报,2013,13(12):89-96.
[37]Zhang X Y,Chen Z X,Wang L. Removal of protein from polysaccharides of rice bran[J]. Science&Technology of Food Industry,2008,29(3):163-165.
[38]Xu H D,Li J,Li Y J,et al.Using macroporous resin AB-8for pigment removal from onion polysaccharides[J].Food Science,2012,33(6):127-131.
[39]付丽红,王艳萍,杨延瑞,等.羊肚菌胞外多糖理化性质及抗氧化活性研究[J].食品科技,2013(11):183-188.
[40]黄俊丽,张慜.松茸、黑牛肝菌、双孢白蘑菇提取物体外抗氧化性试验研究[J].食品与生物技术学报,2011,30(3):342-347.
[41]Zhao B T,Liu J,Chen X,et al.Purification,structure and anti-oxidation of polysaccharides from the fruit of Nitraria tangutorum Bobr[J]. RSC Advances,2018,8(21):11731-11743.
[42]袁红波,张劲松,贾薇,等.利用大孔树脂对低分子量灵芝多糖脱色的研究[J].食品工业科技,2009,30(3):204-206.
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