雪松松针多糖超声波酶法提取及其抗氧化性
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摘要
为优化雪松松针多糖超声波酶法的提取工艺,并研究多糖结构及其抗氧化性。通过响应面法分析确定最佳提取参数为:3. 0 g松针粉末,液料比20∶1(m L∶g),提取温度80℃,超声功率560 W,超声时间47 min,纤维素酶用量12 FPU/g原料,提取两次,多糖得率高达10. 39%。采用高效液相色谱、红外光谱和核磁共振光谱等对松针多糖进行了结构表征,松针多糖以β-糖苷键为主要连接方式,并由葡萄糖、果糖、阿拉伯糖和半乳糖等单糖组成。体外抗氧化性研究结果表明:松针粗多糖对羟基自由基(·OH)和1,1-二苯基-2-三硝基苯肼自由基(DPPH·)的清除能力远高于纯化多糖,呈现出良好的量效关系,粗多糖对·OH和DPPH·的半抑制浓度IC50分别为0. 47 g/L和0. 076 g/L。
In order to optimize the ultrasonic-assisted enzymatic extraction process of polysaccharide from Cedrus deodara pine needles,and investigate the structure of polysaccharide and its antioxidant activity. The optimal extraction parameters were determined by response surface methodology as follows:3. 0 g pine needle powder,liquid to material ratio 20∶ 1(m L∶ g),extraction temperature 80 ℃,ultrasonic power 560 W,ultrasonic extraction time 47 min with the amount of cellulase 12 FPU/g raw material,and extraction twice,the polysaccharide yield was as high as 10. 39%. The structure of pine needle polysaccharide was characterized by high performance liquid chromatography,infrared spectroscopy and nuclear magnetic resonance spectroscopy. The purified polysaccharide,linked mainly by β-glycosidic bonds,consisted of monosaccharides such as glucose,fructose,arabinose and galactose. Antioxidant activities of the polysaccharides were evaluated on the basis of hydroxyl radical(·OH) and 1,1-diphenyl-2-picrylhydrazyl radical(DPPH·). The scavenging ability of the crude polysaccharide was much higher than that of purified polysaccharide,which showed a good dose-effect relationship. The semi-inhibitory concentrations IC50 of the crude polysaccharide on ·OH and DPPH· were 0. 47 g/L and 0. 076 g/L,respectively.
In order to optimize the ultrasonic-assisted enzymatic extraction process of polysaccharide from Cedrus deodara pine needles,and investigate the structure of polysaccharide and its antioxidant activity. The optimal extraction parameters were determined by response surface methodology as follows:3. 0 g pine needle powder,liquid to material ratio 20∶ 1(m L∶ g),extraction temperature 80 ℃,ultrasonic power 560 W,ultrasonic extraction time 47 min with the amount of cellulase 12 FPU/g raw material,and extraction twice,the polysaccharide yield was as high as 10. 39%. The structure of pine needle polysaccharide was characterized by high performance liquid chromatography,infrared spectroscopy and nuclear magnetic resonance spectroscopy. The purified polysaccharide,linked mainly by β-glycosidic bonds,consisted of monosaccharides such as glucose,fructose,arabinose and galactose. Antioxidant activities of the polysaccharides were evaluated on the basis of hydroxyl radical(·OH) and 1,1-diphenyl-2-picrylhydrazyl radical(DPPH·). The scavenging ability of the crude polysaccharide was much higher than that of purified polysaccharide,which showed a good dose-effect relationship. The semi-inhibitory concentrations IC50 of the crude polysaccharide on ·OH and DPPH· were 0. 47 g/L and 0. 076 g/L,respectively.
引文
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2 Zhang X,Sun AD.Study on pine needle extract[J].Food Nutr Chin(中国食物与营养),2009,12(9):23-25.
3 Zhang QL,Gao HQ,Ling LJ,et al.Crude polysaccharides extracted from three characteristic plants in Qinghai province and their activities[J].Nat Prod Res Dev(天然产物研究与开发),2017,29:34-39.
4 Dhayabaran D,Florance EJ,Nandakumar K,et al.Anticonvulsant activity of fraction isolated from ethanolic extract of heartwood of Cedrus deodara[J].J Nat Med,2014,68:310-315.
5 Li PQ,Zhou LG,Mou Y,et al.Extraction optimization of polysaccharide from Zanthoxylum bungeanum using RSM and its antioxidant activity[J].Int J Biol Macromol,2015,72(1):19-27.
6 Yuan M,Jia XJ,Yang Y,et al.Effect of light on structural properties and antioxidant activities of polysaccha-rides from soybean sprouts[J].Process Biochem,2015,50:1152-1157.
7 Ge X,Wang WJ,Xu MS,et al.Extract polysaccharide from cedar pine needles with the method of boiling water-extraction[J].Food Res Dev(食品研究与开发),2010,31(7):20-23.
8 Xu XX,Yin XL,Hong YP,et al.Extraction process and antioxidant activity of Pinus taeda L.polysaccharides[J].Chem Ind Forest Prod(林产化学与工业),2016,36:114-120.
9 Xu XX,Hong YP,Hu JM,et al.Study on microwave extraction technologyand antioxidant activity of pine needle polysaccharose[J].Sci Technol Food Ind(食品工业科技),2015,36:222-227.
10 Niu YY.Effects of polygonatum odoratum polysaccharide on anti-fatigue in mice[J].Nat Prod Res Dev(天然产物研究与开发),2018,30:1202-1207.
11 Yu F,Jiang CY,Yang AP,et al.Study on extraction and purification technology of yam polysaccharides[J].China Condiment(中国调味品),2012,37(12):50-53.
12 Ye M,Wen Z,Wang Q,et al.Effects of deproteinization for dictyophora rubrovalvata polysaccharide by different methods[J].Guizhou Agr Sci(贵州农业科学),2016,44:131-135.
13 Zhou CS,Yu XJ,Zhang YZ,et al.Ultrasonic degradation,purification and analysis of structure and antioxi-dant activity of polysaccharide from Porphyra yezoensis Udea[J].Carbohydr Polym,2012,87:2046-2051.
14 Brummer Y,Cui W,Wang Q.Extraction,purification and physicochemical characterization of fenugreek gum[J].Food Hydrocolloid,2003,17:229-236.
15 Chen HX,Zhang M,Xie BJ.Quantification of uronic acids in tea polysaccharide conjugates and their antiox-idant properties[J].J Agr Food Chem,2004,52:3333-3336.
16 Xie JH,Wang ZJ,Shen MY,et al.Sulfated modification,characterization and antioxidant activities of polysaccharide from Cyclocarya paliurus[J].Food Hydrocolloid,2016,53:7-15.
17 Xie JH,Wang ZJ,Shen MY,et al.Extraction,chemical composition and antioxidant activity of flavonoids from Cyclocarya paliurus(Batal.)Iljinskaja leaves[J].Food Chem,2015,186:97-105.
18 Nie SP,Xie MY.A review on the isolation and structure of tea polysaccharides and their bioactivities[J].Food Hydrocolloid,2011,25:144-149.