Extraction, purification and biological activity of polysaccharide from Glycyrrhiza uralcnsis in Gansu
-
摘要: 以甘肃道地甘草为原料,采用响应面设计法优化超声波辅助热水法提取甘草粗多糖的工艺条件,粗多糖经DEAE-52阴离子交换柱和Sephadex G100凝胶柱纯化获得甘草多糖（GCP2）,利用气相色谱、高效液相色谱联用多角度激光散射技术（HPSEC-LLS）和红外光谱技术对GCP2的单糖组成、分子量分布和光谱特性进行了分析;并评价了GCP2的体外抗氧化性能和对6种供试细菌的抑制活性。结果表明,甘草多糖提取最佳工艺条件:温度为70℃、时间为85 min、液料比为13∶1、超声功率600 W,多糖平均提取得率为4.23%;GCP2是以α-糖苷键为主的还原性多糖,浅黄白色粉末,易溶于水;主要由阿拉伯糖、葡萄糖和半乳糖组成,摩尔比为0.166∶5.56∶1.60。HPSEC-LLS分析结果表明GCP2是由3种不同组分组成的聚合物构成,其中主要组分的重均分子量为1.378×105g/mol,红外光谱显示GCP2具有明显的多糖特征吸收峰,具有α-吡喃糖苷键。甘草多糖对羟自由基和超氧阴离子具有较好的清除能力,活性大小与多糖的浓度呈明显的线性关系,IC50值分别为3.48 mg/m L和3.43 mg/m L,同时还具有一定的还原能力和抑菌作用。实验结果显示,GP2对6种细菌均具有较好的抑制作用,尤其是大肠杆菌和肺炎克雷伯菌。Abstract: Extraction, purification and biological activity of polysaccharide from Glycyrrhiza uralcnsis in Gansu were studied.The optimal ultrasonic-assisted extraction technology of polysaccharide was determined by response surface methodology. The polysaccharide of G. uralensis Fisch. ( GCP2) was obtained by the DEAE-52 anion exchange column chromatography and Sephadex G-100 gel filtration, respectively.The composition and molecular weight of the polysaccharide purified were analyzed by gas chromatography and high performance size exclusion chromatography with multi-angle laser light scattering ( HPSECLLS) .The antioxidant and antibacteria activities of GCP were determined in vitro. Results indicated that the optimal extraction conditions were as follows: extraction temperature 70 ℃, extraction time 85 min, liquid-solid ratio 13 ∶ 1, ultrasonic power600 W. Under the optimal extraction conditions, the extraction rate of polysaccharide was up to 4.23%.GCP2 was obtained by the chromatographic technique, which was a yellow white powder and reducing sugar without starch.GCP2 consists of arabinose, glucose and galactose with the molar ratio of 0.166∶ 5.56∶ 1.60, and had the characteristic of glycoconjugate by infrared spectrum analysis.HPSEC-LLS analysis results showed that the GCP2 was composed of three different components of polymer, average molecular weight of the main components was 1.378 × 10~5g/mol.GCP2 had strong scavenging effect on ·OH free radicals and O2-·free radicals with the IC50 of 3.48 mg/m L and 3.43 mg/m L, and had the reductibility increased with the increase of concentration. Antibacteria test showed that GCP2 had better restraining effect on six different bacteria, especially for Escherichia coli and Pneumonia klebsiella strains.
-
[1] 王巧娥, 任虹, 曹学丽.甘草研究开发与利用现状[J].中国农学通报, 2011, 27 (4) :290-295. [2] Vivek K Gupta, Atiya Fatima, Uzma Faridi, et al.Antimicrobial potential of Glycyrrhiza glabra roots[J].Journal of Ethnopharmacology, 2008, 116 (2) :377-380.
[3] D M Biondi, C Rocco, G Ruberto.New dihydrostibene derivatives from the leaves of Glycyrrhiza glabra and evaluation of their antioxidant activity[J].Journal of Natural Products, 2003, 66 (4) :477-480.
[4] S Demizu, K Kajiyama, K Takahashi, et al.Antioxidant and anti-microbial constituents of licorice:isolation and structure elucidation of a new benzofuran derivative[J].Chemical and Pharmaceutical Bulletin, 1988, 36:3474-3479.
[5] 孙明谦.甘草中化学成分的研究[D].长春:吉林大学, 2006. [6] 常婧.甘草中黄酮类有效成分的提取、含量测定和抗氧化性研究[D].太原:中北大学, 2014. [7] 韩雅慧, 顾赛麒, 陶宁萍, 等.甘草总黄酮提取工艺及总抗氧化活性研究[J].食品工业科技, 2012 (2) :239-242. [8] 张波, 官月平, 田庆来, 等.乌拉尔甘草中黄酮类化学成分的研究进展[J].中成药, 2006, 28 (9) :1362-1365. [9] T Fukai, M Ali, K Kaitou, et al.Anti-Helicobacter pylori flavonoids from licorice extract[J].Life Sciences, 2002, 71 (12) :1449-1463.
[10] T Fukai, A Marumo, K Kaitou, et al.Antimicrobial activity of licorice flavonoids against methicillin-resistant Staphylococcus aureus[J].Fitoterapia, 2002, 73 (6) :536-539.
[11] 田庆来, 官月平, 张波, 等.甘草有效成分的药理作用研究进展[J].天然产物研究与开发, 2006, 18 (2) :343-347. [12] 聂小华, 尹光耀, 史宝军, 等.甘草有效成分体外抗肿瘤活性和免疫活性的研究[J].中药材, 2003, 26 (7) :507-509. [13] H Ying Kai, W Hua Tao, M Tao, et al.Effects of Glycyrrhiza glabra polysaccharides on immune and antioxidant activities in high-fat mice[J].International Journal of Biological Macromolecules, 2009, 45 (1) :61-64.
[14] 张泽生, 史珅, 白鑫, 等.新疆产甘草品质评价及甘草多糖提取工艺研究[J].现代食品科技, 2008, 24 (2) :143-146. [15] 刘文玉, 颜雪琴, 林祥群, 等.响应面法优化新疆胀果甘草多糖的提取工艺[J].食品工业科技, 2012, 33 (23) :288-291. [16] 代巧玉, 康丁, 张洪斌.水溶性甘草多糖的分离纯化、结构及生物活性研究进展[J].天然产物研究与开发, 2013, 25 (9) :1292-1296. [17] 张琳, 樊金玲, 朱文学, 等.响应面法优化超声波辅助提取甘草多糖工艺[J].食品科学, 2010, 16:67-71. [18] 王忱, 谢广茹, 史玉荣, 等.甘草多糖的体内抑瘤作用及其机制的研究[J].临床肿瘤学杂志, 2003, 8 (2) :85-87. [19] 张泽生, 史珅, 杨超慧, 等.甘草多糖免疫调节作用的研究[J].现代生物医学进展, 2008, 8 (10) :1835-1837. [20] 王忱.甘草多糖抑瘤的体内实验及其分子机制的研究[D].天津:天津医科大学, 2002. [21] 热娜·卡斯木, 丛媛媛, 屠鹏飞.胀果甘草多糖的分离纯化及其理化性质[J].华西药学杂志, 2008, 23 (4) :448-450. [22] 刘晓风, 刘琳, 王永刚, 等.啤特果多糖分离纯化及抗氧化活性研究[J].现代食品科技, 2014, 30 (7) :179-186. [23] 刘建利.宁夏乌拉尔甘草多糖的提取及其体外抗氧化活性的测定[J].食品与发酵工业, 2011, 37 (7) :210-213. [24] L Wei, W Heng Yu, P Xiu Bing, et al.Characterization and antioxidant activity of two low-molecular-weight polysaccharides purified from the fruiting bodies of Ganoderma lucidum[J].International Journal of Biological Macromolecules, 2010, 46 (4) :451-457.
[25] 张百刚, 钟旭美, 刘晓风, 等.甘草多糖的提取及其抑菌实验的研究[J].粮油加工, 2010 (8) :137-140. [26] 孙润广, 张静.甘草多糖螺旋结构的原子力显微镜研究[J].化学学报, 2006, 24 (64) :2467-2472. [27] 周蓉, 齐莉, 王雅芬, 等.甘草多糖的分离纯化及高效毛细管电泳分析[J].分析化学, 1999, 27 (2) :245. [28] 艾则孜, 马小春.HPLC-ELSD法分析甘草多糖中单糖的组成[J].新疆大学学报 (自然科学版) , 2007, 24:236-238. [29] 张惟杰.糖复合物生化研究技术[M].浙江:浙江大学出版社, 1999.
计量
- 文章访问数: 216
- HTML全文浏览量: 22
- PDF下载量: 780
下载:
