牛樟芝总三萜超声提取及体外活性研究
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摘要
为有效利用牛樟芝资源,以人工固体培养牛樟芝菌丝体为原料,采用响应面法优化牛樟芝总三萜超声辅助提取工艺,比色法评价牛樟芝总三萜体外抗氧化活性,滤纸片法研究牛樟芝总三萜的体外抑菌活性。结果表明,超声辅助提取牛樟芝总三萜的最佳工艺条件为:提取时间41 min、提取功率80 W、料液比1∶20,总三萜得率为3.84%±0.18%。牛樟芝三萜提取物具有较强的还原能力,并能有效清除DPPH自由基、ABTS自由基和超氧阴离子自由基,其半清除浓度(EC_(50))分别为0.29、0.50、0.33mg·m L~(-1),且清除率与质量浓度存在剂量效应关系;牛樟芝总三萜对大肠杆菌、金黄色葡萄球菌及枯草芽孢杆菌均具有明显的抑制作用,同浓度下对金黄色葡萄球菌具有较高的抑菌活性。本研究结果为人工固体栽培牛樟芝活性成分的综合开发利用提供了依据。
In order to effectively utilize the resource of Antrodia camphorata, the extraction conditions of total triterpenoids from mycelium of solid-state cultured Antrodia camphorata were optimized by response surface analysis( RSE). In vitro antioxidant activities of triterpenoids were evaluated by colorimetry. Antibacterial activities of triterpenoids were determined by filtering paper method. The results showed that the optimum ultrasonic-assisted extraction conditions were as follows: extraction time of 41 min,extraction power of 80 W,and a ratio of solid to liquid as 1: 20. And the yield of total triterpenoids from Antrodia camphorata was 3. 84% ± 0. 18%. The iron ions reduction results showed that triterpenoids had certain reducing power. The EC_(50) values of free radical scavenging capacities of DPPH,ABTS and superoxide anions was 0. 29,0. 50,and 0. 33 mg·m L~(-1),respectively. The triterpenoids have strong inhibition effects against Escherichia coli,Staphylococcus aureus and Bacillus subtilis. Higher antibacterial activity was found to be against of Staphylococcus aureus when used at the same concentration of triterpenoids. In conclusion,total triterpenoids extracts from Antrodia camphorata showed relatively strong antioxidant activities and antibacterial activities.This work could provide useful information for further development and comprehensive utilization of total triterpenoids from solid-state cultured Antrodia camphorata.
In order to effectively utilize the resource of Antrodia camphorata, the extraction conditions of total triterpenoids from mycelium of solid-state cultured Antrodia camphorata were optimized by response surface analysis( RSE). In vitro antioxidant activities of triterpenoids were evaluated by colorimetry. Antibacterial activities of triterpenoids were determined by filtering paper method. The results showed that the optimum ultrasonic-assisted extraction conditions were as follows: extraction time of 41 min,extraction power of 80 W,and a ratio of solid to liquid as 1: 20. And the yield of total triterpenoids from Antrodia camphorata was 3. 84% ± 0. 18%. The iron ions reduction results showed that triterpenoids had certain reducing power. The EC_(50) values of free radical scavenging capacities of DPPH,ABTS and superoxide anions was 0. 29,0. 50,and 0. 33 mg·m L~(-1),respectively. The triterpenoids have strong inhibition effects against Escherichia coli,Staphylococcus aureus and Bacillus subtilis. Higher antibacterial activity was found to be against of Staphylococcus aureus when used at the same concentration of triterpenoids. In conclusion,total triterpenoids extracts from Antrodia camphorata showed relatively strong antioxidant activities and antibacterial activities.This work could provide useful information for further development and comprehensive utilization of total triterpenoids from solid-state cultured Antrodia camphorata.
引文
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[25]张寅,许泓瑜,陆震鸣,张晓梅,窦文芳,许正宏.薄层色谱-分光光度法测定樟芝菌粉中的总三萜含量[J].天然产物研究与开发,2010,22(4):639-642
[26]陆震鸣,陶文沂,许泓瑜,敖宗华,许正宏.樟芝菌粉三萜类化合物含量的测定[J].中成药,2008,30(3):402-405
[27]周晓,王成忠,李双,王晓华.响应面法优化超声辅助提取灵芝三萜的工艺研究[J].食品工业,2015,36(6):136-140
[28]Yang F C,Yang Y H,Lu H C.Enhanced antioxidant and antitumor activities of Antrodia cinnamomea,cultured with cereal substrates in solid state fermentation[J].Biochemical Engineering Journal,2013,78(37):108-113
[29]汪艳群,孟宪军.超声波处理对北五味子多糖抗氧化活性的影响[J].食品科学,2016,37(3):66-70
[30]Amoussa A M O,Lagnika L,Bourjot M,Vonthron-Senecheau C,Sanni A.Triterpenoids from Acacia ataxacantha DC:antimicrobial and antioxidant activities[J].BMC Complementary and Alternative Medicine,2016,16(1):284-292
[31]孙延芳,梁宗锁,刘政,黄静,闫晗.酸枣果三萜皂苷抑菌和抗氧化活性的研究[J].食品工业科技,2012,33(6):139-142
[2]徐小伟,泽桑梓,杨斌,季梅.牛樟芝主要活性成分及其药理研究现状[J].林业调查规划,2014,39(6):21-25
[3]韩金龙,赵培城,毛建卫,刘士旺.樟芝发酵液和菌丝体提取物抗氧化性能研究[J].微生物学杂志,2014,34(5):49-54
[4]Mau J L,Huang P N,Huang S J,Chen C C.Antioxidant properties of methanolic extracts from two kinds of Antrodia camphorata mycelia[J].Food Chemistry,2004,86(1):25-31
[5]Lien H M,Tseng C J,Huang C L,Lin Y T,Chen C C,La Y Y.Antimicrobial Activity of Antrodia camphorata extracts against oral bacteria[J].PLo S One,2014,9(8):105-112
[6]李晔,朱忠敏,姚渭溪,陈若芸.灵芝三萜类化合物的研究进展[J].中国中药杂志,2012,37(2):165-171
[7]Hossain M B,Tiwari B K,Gangopadhyay N,Donnell C P O,Brunton N P,Rai D K.Ultrasonic extraction of steroidal alkaloids from potato peel waste[J].Ultrasonics Sonochemistry,2014,21(4):1470-1476
[8]Schinor E C,Salvador M J,Turatti I C,Zucchi O L.Comparison of classical and ultrasound-assisted extractions of steroids and triterpenoids from three Chresta spp[J].Ultrasonics Sonochemistry,2004,11(6):415-421
[9]郭丹钊,马海乐,曹颖王,王文秀,王亚,王薇薇.桑黄菌丝体活性物质的超声波辅助提取工艺[J].中国食品学报,2015,15(4):87-92
[10]庾石山.三萜化学[M].北京:化学工业出版社,2008:74
[11]Heleno S A,Barros L,Martins A,Mari J R Q,Morales P,Virgini F R,Isabel C F.Chemical composition,antioxidant activity and bioaccessibility studies in phenolic extracts of two Hericium wild edible species[J].LWT-Food Science and Technology,2015,63(1):475-481
[12]Qiu J,Wang X,Song C.Neuroprotective and antioxidant lanostanoid triterpenes from the fruiting bodies of Ganoderma atrum[J].Fitoterapia,2016,109:75-79
[13]Li B,Zhang X Y,Wang M Z,Jiao L L.Characterization and antioxidant activities of acidic polysaccharides from Gynostemma pentaphyllum(Thunb.)Markino[J].Carbohydrate Polymers,2015,127:209-214
[14]Singh P,Singh R.,Sati N,Sati O P,Kumar N.Antioxidant and antibacterial activity of Jurinea dolomiaea Boiss extracts[J].International Journal of Life-Sciences Scientific Research,2015,1(2):74-78
[15]Feng K,Chen W,Sun L W,Liu J Z,Zhao Y X,Li L X,Wang Y X,Zhang W J.Optimization extraction,preliminary characterization and antioxidant activity in vitro of polysaccharides from Stachys sieboldii Miq.tubers[J].Carbohydrate Polymers,2015,125:45-52
[16]赖谱富,赖锋连,陈君琛,沈恒胜.多年生藤本豆总黄酮纯化及抑菌活性研究[J].核农学报,2015,29(8):1539-1546
[17]Awanchiri S S,Trinhvandufat H,Shirri J C,Dongfack M D J,Nguenang G M.Boutefnouchet S,Fomum Z T,Seguin E,Verite P,Tillequin F,Wandji J.Triterpenoids with antimicrobial activity from Drypetes inaequalis[J].Phytochemistry,2009,70(3):419-423
[18]刘慧,张春岭,陈大磊,李佳秀,刘杰超,焦中高.响应面法优化超声波提取山楂皮渣三萜酸工艺[J].保鲜与加工,2017,17(1):53-59
[19]Maran J P,Mekala V,Manikandan S.Modeling and optimization of ultrasound-assisted extraction of polysaccharide from Cucurbita moschata[J].Carbohydrate Polymers,2013,92(2):2018-2026
[20]Wang J,Sun B G,Cao Y P,Tian Y,Li X H.Optimisation of ultrasound-assisted extraction of phenolic compounds from wheat bran[J].Food Chemistry,2008,106(2):804-810
[21]李明华,陆正清,孟秀梅,陈军.金针菇多糖闪式提取工艺及其抗氧化活性研究[J].食品与发酵工业,2016,42(10):216-221
[22]Geethangili M,Tzeng Y M.Review of pharmacological effects of Antrodia camphorata and its bioactive compounds[J].Evidencebased Complementary and Alternative Medicine,2011:1-17
[23]王蓓,王薇薇,马海乐,叶晓非.静磁场辅助液态发酵对牛樟芝菌丝体及三萜产量的影响[J].中国农业科技导报,2015,17(5):99-105
[24]弓威,顾丰颖,贺凡,王锋.山楂叶有效成分提取工艺优化及抗氧化活性研究[J].核农学报,2015,29(8):1547-1558
[25]张寅,许泓瑜,陆震鸣,张晓梅,窦文芳,许正宏.薄层色谱-分光光度法测定樟芝菌粉中的总三萜含量[J].天然产物研究与开发,2010,22(4):639-642
[26]陆震鸣,陶文沂,许泓瑜,敖宗华,许正宏.樟芝菌粉三萜类化合物含量的测定[J].中成药,2008,30(3):402-405
[27]周晓,王成忠,李双,王晓华.响应面法优化超声辅助提取灵芝三萜的工艺研究[J].食品工业,2015,36(6):136-140
[28]Yang F C,Yang Y H,Lu H C.Enhanced antioxidant and antitumor activities of Antrodia cinnamomea,cultured with cereal substrates in solid state fermentation[J].Biochemical Engineering Journal,2013,78(37):108-113
[29]汪艳群,孟宪军.超声波处理对北五味子多糖抗氧化活性的影响[J].食品科学,2016,37(3):66-70
[30]Amoussa A M O,Lagnika L,Bourjot M,Vonthron-Senecheau C,Sanni A.Triterpenoids from Acacia ataxacantha DC:antimicrobial and antioxidant activities[J].BMC Complementary and Alternative Medicine,2016,16(1):284-292
[31]孙延芳,梁宗锁,刘政,黄静,闫晗.酸枣果三萜皂苷抑菌和抗氧化活性的研究[J].食品工业科技,2012,33(6):139-142