麻栎多糖的提取、纯化及其性质分析
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摘要
为了开发利用麻栎多糖资源,探讨了热水浸提法提取麻栎多糖的工艺参数并对其纯多糖进行了初步分析。以麻栎多糖的得率为指标,通过单因素试验对提取时间、温度、液料比和提取次数进行考察,进一步通过响应面分析法进行优化,得到提取多糖的最佳条件为:提取时间3 h,提取温度90℃,液料比25∶1(mL∶g),提取次数2次,在此条件下多糖得率为2.65%。麻栎粗多糖经DEAE-52离子交换柱和Sephadex G-200凝胶柱两步分离纯化后,得到2种纯多糖OP-1和OP-2。紫外和红外光谱分析结果表明:OP-1和OP-2的纯度较高,具有多糖的特征吸收峰;经柱前衍生高效液相色谱法测定结果表明:OP-1是由木糖和葡萄糖组成的中性杂多糖,其物质的量之比为0.86∶0.7;OP-2是由核糖、葡萄糖醛酸和阿拉伯糖组成的酸性杂多糖,其物质的量之比为0.42∶0.21∶0.56;采用高效凝胶渗透色谱(HPGPC-ELSD)法测得OP-1的重均分子质量为444.8 ku,OP-2的重均分子质量为121.2 ku。
In order to develop and utilize the polysaccharide from Quercus acutissima, the process parameters of hot water extraction of polysaccharides were studied and the purified polysaccharide was preliminarily analyzed. The extraction conditions including extraction time, temperature, the liquid to solid ratio and times of extraction were investigated by single factor experiment and response surface method with the yield of polysaccharides as index. The optimal extraction conditions were the extraction time 3 h, the extraction temperature 90 ℃, the liquid to solid ratio 25∶1(mL∶g), and the times of extraction 2. Under these conditions, polysaccharide yield was 2.65%. By using DEAE-52 and Sephadex G-200 column chromatography, two purified polysaccharides(OP-1, OP-2) were obtained. The results of ultraviolet and FT-IR spactra, showed that the purified OP-1 and OP-2 from Q. acutissima had high purity and the typical groups of polysaccharide. The composition analysis by HPLC with pre-column derivatization indicated that the OP-1, a neutral heteropolysaccharide, was mainly composed of xylose and glucose with the molar ratio of 0.86∶0.7; the OP-2, an acidic heteropolysaccharide, was mainly composed of ribose, glucuronic acid, and arabinose with the molar ratio of 0.42∶0.21∶0.56. The results of high performance gel permeation chromatography(HPGPC-ELSD) indicated that the weight average molecular weights(M_w) of OP-1 and OP-2 were about 444.8 and 121.2 ku.
In order to develop and utilize the polysaccharide from Quercus acutissima, the process parameters of hot water extraction of polysaccharides were studied and the purified polysaccharide was preliminarily analyzed. The extraction conditions including extraction time, temperature, the liquid to solid ratio and times of extraction were investigated by single factor experiment and response surface method with the yield of polysaccharides as index. The optimal extraction conditions were the extraction time 3 h, the extraction temperature 90 ℃, the liquid to solid ratio 25∶1(mL∶g), and the times of extraction 2. Under these conditions, polysaccharide yield was 2.65%. By using DEAE-52 and Sephadex G-200 column chromatography, two purified polysaccharides(OP-1, OP-2) were obtained. The results of ultraviolet and FT-IR spactra, showed that the purified OP-1 and OP-2 from Q. acutissima had high purity and the typical groups of polysaccharide. The composition analysis by HPLC with pre-column derivatization indicated that the OP-1, a neutral heteropolysaccharide, was mainly composed of xylose and glucose with the molar ratio of 0.86∶0.7; the OP-2, an acidic heteropolysaccharide, was mainly composed of ribose, glucuronic acid, and arabinose with the molar ratio of 0.42∶0.21∶0.56. The results of high performance gel permeation chromatography(HPGPC-ELSD) indicated that the weight average molecular weights(M_w) of OP-1 and OP-2 were about 444.8 and 121.2 ku.
引文
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[22]王丽波,程龙,徐雅琴,等.南瓜籽多糖热水提取工艺优化及其抗氧化活性[J].农业工程学报,2016,32(9):284-290.WANG L B,CHENG L,XU Y Q,et al.Extraction technology optimization and antioxidant activity of polysaccharides from pumpkin seeds[J].Transactions of the Chinese Society of Agricultural Engineering,2016,32(9):284-290.
[23]YU Z Y,LIU L,XU Y Q,et al.Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits[J].Carbohydrate Polymers,2015,132(68):180-186.
[2]陈京元,王义勋,徐红梅,等.刺槐香菇与麻栎香菇的质量比较[J].中南民族大学学报:自然科学版,2017,36(1):57-60.CHEN J Y,WANG Y X,XU H M,et al.The quality comparison of letinous edodes and Quercus acacia letinous edodes [J].Journal of South-Central University for Nationalities:Natural Science Edition,2017,36 (1):57-60.
[3]王义勋,林亲雄,查玉平,等.枫香与栎木基质栽培香菇比较分析[J].湖北农业科学,2016,55(22):5854-5857.WANG Y X,LIN Q X,CHA Y P,et al.The comparative analysis of Lentinus edodes cultivated with Liquidambar formosana and oak wood substrate[J].Hubei Agricultural Science,2016,55(22):5854-5857.
[4]刘志龙,虞木奎,唐罗忠,等.麻栎资源研究进展及开发利用对策[J].中国林副特产,2009(6):93-96.LIU Z L,YU M K,TANG L Z,et al.Progress and utilization countermeasures of Quercus acutissima [J].Forest By-Product and Speciality in China,2009(6):93-96.
[5]胡惠萍,周洁莹,夏凤娜,等.不同原料栽培的灵芝孢子粉质量比较试验[J].食用菌,2008,30(4):27-29.HU H P,ZHOU J Y,XIA F N,et al.The quality comparison test of Ganoderma lucidum spore powder for different raw materials [J].Edible Fungi,2008,30(4):27-29.
[6]刘荣松,李朝谦.六个阔叶树种对栽培灵芝孢子粉产量和质量的影响试验[J].浙江食用菌,2008,16(3):38,45.LIU R S,LI C Q.Effects of six broad-leaved tree species on the yield and quality of cultivated Ganoderma lucidum spore powder [J].Zhejiang Shiyongjun,2008,16(3):38,45.
[7]鲍文辉.不同栽培基质培养的灵芝子实体品质差异性研究[J].食药用菌,2015,23(5):318-319.BAO W H.Study on quality difference of Ganoderma lucidum fruiting bodies cultured in different culture substrates [J].Edible and Medicinal Mushrooms,2015,23(5):318-319.
[8]周磊,许敏,杨崇仁,等.壳斗科植物的化学成分及生物活性研究进展[J].天然产物研究与开发,2012,24(2):260-273.ZHOU L,XU M,YANG C R,et al.The advances of chemical components and bioactivity of Fagaceae plants [J].Natural Product Research and Development,2012,24(2):260-273.
[9]LUO L,ZHENG S S,HUANG Y F,et al.Preparation and characterization of Chinese yam polysaccharide PLGA nanoparticles and their immunological activity[J].International Journal of Pharmaceutics,2016,511(1):140-150.
[10]LI X,LIANG J.Ganoderma lucidum polysaccharide prevents oxidation and skin aging[J].Chinese Journal of Tissue Engineering Research,2013,17(41):7272-7277.
[11]JU Y,XUE Y,HUANG J L,et al.Antioxidant Chinese yam polysaccharides and its pro-proliferative effect on endometrial epithelial cells[J].International Journal of Biological Macromolecules,2014,66(5):81-85.
[12]JIANG J H,SLIVOVA V,HARVEY K,et al.Ganoderma lucidum suppress growth of breast cancer cells through the inhibition of Akt/NF-kappa B signaling[J].Nutrition Cancer,2004,49(2):209-216.
[13]董汝晶.多糖提取方法的研究进展[J].农产品加工(学刊),2014(4):46-48.DONG R J.The research progress of polysaccharide extraction method [J].Academic Periodical of Farm Products Processing,2014(4):46-48.
[14]蒋水星,陈雪峰,赵天殊.响应面法优化大枣多糖的提取工艺研究[J].安徽农业科学,2011,39(6):3652-3654.JIANG S X,CHEN X F,ZHAO T S.Study of optimization of extraction technology of jujube polysaccharides by response surface methodology [J].Journal of Anhui Agricultural Sciences,2011,39(6):3652-3654.
[15]程旺开,许月明,张冬冬.响应面优化黄秋葵叶多糖的提取工艺及其抗氧化活性考察[J].中国实验方剂学杂志,2017,23(4):38-42.CHENG W K,XU Y M,ZHANG D D.Response surface methodology for optimization of extraction and antioxidant activities of polysaccharides from Abelmoschus esculentus leaves [J].Chinese Journal of Experimental Tranditional Medical Formulae,2017,23(4):38-42.
[16]张惟杰.糖复合物生化研究技术[M].杭州:浙江大学出版社,1999.ZHANG W J.Biochemistry Technology of Glycocomplex [M].Hangzhou:Zhejiang University Press,1999.
[17]王向红,吉爽爽,生庆海.柱前衍生高效液相色谱法检测8种枣水溶性多糖的单糖组成[J].中国食品学报,2014,14(9):257-262.WANG X H,JI S S,SHENG Q H.Determination of monosaccharide components of eight varieties jujube water soluble polysaccharides by pre-column derivatization HPLC method [J].Journal of Chinese Istitute of Food Science and Technologe,2014,14 (9):257-262.
[18]许杜娟,夏泉,刘钢,等.高效液相法测定黄芪中性多糖的单糖组成[J].中国实验方剂学杂志,2008,14(9):4-6.XU D J,XIA Q,LIU G,et al.Analysis of monosaccharide composition in Astragalus neutral polysaccharides by high performance liquid chromatography[J].Chinese Journal of Experimental Tranditional Medical Formulae,2008,14(9):4-6.
[19]王川,张燕,李楠,等.HPSEC-ELSD法同时测定白及多糖的分子量和含量[J].中国药房,2018,29(9):1198-1201.WANG C,ZHANG Y,LI N,et al.Determination of molecular weight and content of Bletilla striata polysaccharide by HPSEC-ELSD [J].China Pharmacy,2018,29(9):1198-1201.
[20]圣志存,郝利民,贾士儒,等.响应面法优化红缘拟层孔菌胞外多糖发酵培养基[J].食品工业科技,2014,35(4):194-198.SHENG Z C,HAO L M,JIA S R,et al.Optimization of culture medium of Fomitopsis pinicola for exopolysaccharide production by response surface methodology[J].Science and Technology of Food Industry,2014,35 (4):194-198.
[21]曾红亮,卢旭,卞贞玉,等.响应面分析法优化金柑多糖的提取工艺[J].福建农林大学学报:自然科学版,2012,41(3):315-319.ZENG H L,LU X,BIAN Z Y,et al.Optimization of the extraction technique of Fortunella margarita polysaccharides via response surface analysis [J].Journal of Fujian Agriculture and Forestry University:Natural Science Edition,2012,41(3):315-319.
[22]王丽波,程龙,徐雅琴,等.南瓜籽多糖热水提取工艺优化及其抗氧化活性[J].农业工程学报,2016,32(9):284-290.WANG L B,CHENG L,XU Y Q,et al.Extraction technology optimization and antioxidant activity of polysaccharides from pumpkin seeds[J].Transactions of the Chinese Society of Agricultural Engineering,2016,32(9):284-290.
[23]YU Z Y,LIU L,XU Y Q,et al.Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits[J].Carbohydrate Polymers,2015,132(68):180-186.