辣木多糖酶促提取技术优化及不同部位含量研究
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摘要
为研究辣木多糖的酶促提取工艺和辣木不同部位的多糖含量,在单因素试验的基础上,以多糖提取率为指标,选择纤维素酶用量、提取温度、提取时间、液料比为试验因素,采用二次回归正交旋转组合设计对纤维素酶辅助提取辣木多糖的工艺参数进行优化,并在此基础上研究辣木不同部位的多糖含量差异。4种因素对辣木多糖提取率的影响顺序依次为:纤维素酶用量>提取温度>液料比>提取时间。建立辣木多糖提取率(Y)与纤维素酶用量(X1)、提取温度(X2)、提取时间(X3)、液料比(X4)的二次正交回归模型:Y=18.6602+0.8134X1+0.7572X2–0.4312X3+0.6909X4–0.5181X12–0.4935X22–0.6277X42,该模型拟合度好,4个因素均对多糖提取率有显著影响(p<0.05)。通过回归模型获得优化的提取工艺为:纤维素酶用量1.60%,提取温度53℃,提取时间68 min,液料比52∶1,在此条件下辣木多糖提取率为19.83%,实际值与预测值一致。辣木不同部位的多糖含量结果表明,辣木根、花、嫩叶、茎中均含有丰富的多糖,其中根中含量最高且极显著高于其他部位(p<0.01),可进一步开发利用。采用二次回归正交旋转组合设计优化得到的辣木多糖酶促提取工艺条件准确可靠,可有效提高辣木多糖提取率并可在生产中推广应用。
Single factor experiments incluing enzymatic dosage, extraction temperature, extraction time, ratio of liquid to material, and quadratic regression orthogonal rotation combination design were used to optimize the enzymatic extraction process of polysaccharide from Moringa oleifera Lam.. The quadratic orthogonal regression model of extracted polysaccharide rate(Y) for enzymatic dosage(X1), extraction temperature(X2), extraction time(X3) and ratio of liquid to material(X4) was Y=18.6602+0.8134X1+0.7572X2–0.4312X3+0.6909X4–0.5181X12–0.4935X22–0.6277X42, which had a good fitting degree. The four factors had significant effects on rate of polysaccharide extracted(p<0.05). Furthermore, the optimal compound enzymatic extraction process conditions were as follows: enzymatic dosage of 1.60%, extraction temperature of 53 ℃, extraction time of 68 min, and ratio of liquid to material of 52∶1. Under optimal conditions, the verified value of extracted polysaccharide rate was 19.83%. The contents of polysaccharide from different parts of Moringa had remarkable difference. The content was ranked downward in root, flower, tender leaves, and stalk. It was accurate and reliable that the quadratic regression orthogonal rotation combination design was used to optimize enzymatic extraction process conditions,and the extraction efficiency of polysaccharide from Moringa could be improved effectively, so it could be applied in the actual production. Furthermore, the content of polysaccharide in the root of Moringa was the highest and it was worthy of exploitation and utilization.
Single factor experiments incluing enzymatic dosage, extraction temperature, extraction time, ratio of liquid to material, and quadratic regression orthogonal rotation combination design were used to optimize the enzymatic extraction process of polysaccharide from Moringa oleifera Lam.. The quadratic orthogonal regression model of extracted polysaccharide rate(Y) for enzymatic dosage(X1), extraction temperature(X2), extraction time(X3) and ratio of liquid to material(X4) was Y=18.6602+0.8134X1+0.7572X2–0.4312X3+0.6909X4–0.5181X12–0.4935X22–0.6277X42, which had a good fitting degree. The four factors had significant effects on rate of polysaccharide extracted(p<0.05). Furthermore, the optimal compound enzymatic extraction process conditions were as follows: enzymatic dosage of 1.60%, extraction temperature of 53 ℃, extraction time of 68 min, and ratio of liquid to material of 52∶1. Under optimal conditions, the verified value of extracted polysaccharide rate was 19.83%. The contents of polysaccharide from different parts of Moringa had remarkable difference. The content was ranked downward in root, flower, tender leaves, and stalk. It was accurate and reliable that the quadratic regression orthogonal rotation combination design was used to optimize enzymatic extraction process conditions,and the extraction efficiency of polysaccharide from Moringa could be improved effectively, so it could be applied in the actual production. Furthermore, the content of polysaccharide in the root of Moringa was the highest and it was worthy of exploitation and utilization.
引文
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[20]王晓红,孙延龙,李万忠,等.产地和生长年限对银杏叶中多糖和黄酮含量的影响[J].中药材,2016,39(6):1341-1342.
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[22]王建华,王汉中,张民,等.枸杞多糖延缓衰老的作用[J].营养学报,2002(2):189-191,194.
[23]Jiang B,Zhang H Y,Liu C J,et al.Extraction of water-soluble polysaccharide and the antioxidant activity from Ginkgo biloba leaves[J].Medicinal Chemistry Research,2010,19(3):262-270.
[24]Pillai T G,Nair C K,Janardhanan K K,et al.Polysaccharides isolated from Ganoderma lucidum occurring in Southern parts of India,protects radiation induced damages both in vitro and in vivo[J].Environmental Toxicology&Pharmacology,2008,26(1):80-85.
[25]华允芬.铁皮石斛多糖成分研究[D].杭州:浙江大学,2005.
[26]付立忠,吴学谦,李明焱,等.灵芝品种子实体多糖和三萜含量分析与评价[J].中国食用菌,2009,28(4):38-40.
[27]吴涛,庞坤宇,侯杰,等.枸杞多糖含量种间差异比较[J].北华大学学报(自然科学版),2013,14(5):594-596.
[28]陈开娟.不同等级枸杞中枸杞多糖的含量测定与比较[J].海峡药学,2011,23(8):100-102.
[29]姜波,王艳颖,刘长建,等.银杏叶多糖提取、纯化及其含量的测定[J].时珍国医国药,2007,18(11):2729-2731.
[2]张幸怡,李洋,林聪,等.辣木叶粉对大鼠生长性能、血液与肝脏抗氧化及免疫指标的影响[J].天然产物研究与开发,2016,28(11):1724-1731.
[3]Jung I L.Soluble extract from Moringa oleifera leaves with a new anticancer activity[J].PLoS One,2014,9(4):95492.
[4]Marrufo T,Nazzaro F,Mancini E,et al.Chemical composition and biological activity of the essential oil from leaves of Moringa oleifera Lam.cultivated in Mozambique[J].Molecules,2013,18(9):10989-11000.
[5]Maldini M,Maksoud S A,Natella F,et al.Moringa oleifera:study of phenolics and glucosinolates by mass spectrometry[J].Journal of Mass Spectrometry,2014,49(9):900.
[6]Brekovich L,Earon G,Ron I,et al.Moringa oleifera aqueous leaf extract down-regulates nuclear factor-kappa B and increases cytotoxic effect of chemotherapy in pancreatic cancer cells[J].BMC Complementary and Alternative Medicine,2013,13(1):212.
[7]Murakami A,Kitazono Y,Jiwajinda S,et al.Niaziminin,a thiocarbamatc from the leaves of Moringa oleifera,holds a strict structure requirement for inhibition of tumor-promoter-indccd Epstein-Barr virus activation[J].Planta Medica,1998,64(4):319-323.
[8]曹玉霖,何镇廷,朱彦锋.辣木的抗肿瘤活性及作用机制研究进展[J].天然产物研究与开发,2016,28(11):1845-1849.
[9]梁鹏,甄润英.辣木茎叶中水溶性多糖的提取及抗氧化活性的研究[J].食品研究与开发,2013,34(14):25-29.
[10]杜洋.辣木多糖的提取与体外抗丙肝病毒活性的研究[D].成都:四川农业大学,2015.
[11]石奇.植物多糖的新型提取分离技术应用进展[J].西安文理学院学报(自然科学版),2015,18(3):50-54.
[12]彭凌.辣木多糖超声波辅助提取工艺条件优化研究[J].江苏农业科学,2008(4):238-239.
[13]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):282-286.
[14]陈瑞娇.辣木多糖的提取及分离纯化[J].中草药,2006,29(12):1358-1360.
[15]李绍平,黄赵刚,张平,等.蒽酮-硫酸法测定亮菌糖浆中多糖的含量[J].中草药,2002,33(3):233-234.
[16]李云雁,胡传荣.试验设计与数据处理[M].第2版,北京:化学工业出版社,2008.
[17]孙鸣燕.辣木黄酮和多糖提取方式及其含量影响因素的初步研究[D].呼和浩特:内蒙古农业大学,2008.
[18]徐金楠,刘玮,刘春晶,等.不同枸杞中多糖含量与结构特征的对比研究[J].中国食品学报,2015,15(4):233-239.
[19]谢玉策,柯乐芹,张松成.香菇多糖含量的影响因素[J].浙江农业科学,2013,1(12):1632-1633.
[20]王晓红,孙延龙,李万忠,等.产地和生长年限对银杏叶中多糖和黄酮含量的影响[J].中药材,2016,39(6):1341-1342.
[21]Meng L Z,Lv G P,Hu D J,et al.Effects of polysaccharides from different species of Dendrobium(Shihu)on macrophage function[J].Molecules,2013,18(5):5779-5791.
[22]王建华,王汉中,张民,等.枸杞多糖延缓衰老的作用[J].营养学报,2002(2):189-191,194.
[23]Jiang B,Zhang H Y,Liu C J,et al.Extraction of water-soluble polysaccharide and the antioxidant activity from Ginkgo biloba leaves[J].Medicinal Chemistry Research,2010,19(3):262-270.
[24]Pillai T G,Nair C K,Janardhanan K K,et al.Polysaccharides isolated from Ganoderma lucidum occurring in Southern parts of India,protects radiation induced damages both in vitro and in vivo[J].Environmental Toxicology&Pharmacology,2008,26(1):80-85.
[25]华允芬.铁皮石斛多糖成分研究[D].杭州:浙江大学,2005.
[26]付立忠,吴学谦,李明焱,等.灵芝品种子实体多糖和三萜含量分析与评价[J].中国食用菌,2009,28(4):38-40.
[27]吴涛,庞坤宇,侯杰,等.枸杞多糖含量种间差异比较[J].北华大学学报(自然科学版),2013,14(5):594-596.
[28]陈开娟.不同等级枸杞中枸杞多糖的含量测定与比较[J].海峡药学,2011,23(8):100-102.
[29]姜波,王艳颖,刘长建,等.银杏叶多糖提取、纯化及其含量的测定[J].时珍国医国药,2007,18(11):2729-2731.