蛹拟青霉发酵液中抑菌活性成分的分离纯化
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摘要
以大肠杆菌、白色假丝酵母、黑曲霉为指示菌株,对蛹拟青霉xzcs005菌株液体发酵液乙酸乙酯萃取相中的抑菌活性成分进行分离纯化。乙酸乙酯萃取相经硅胶柱色谱分离后得到组分Ⅴ和组分Ⅶ两个抑菌活性组分。组分Ⅴ利用反相柱分离得到抑菌活性纯品PMBA-1,其在质量浓度为1.0mg/mL时对大肠杆菌、白色假丝酵母和黑曲霉的抑菌圈直径分别为(16.21±0.27)、(23.54±0.35)、(27.33±0.54)mm;组分Ⅶ再次经过硅胶柱色谱分离后得到抑菌活性纯品PMBA-2,其在质量浓度为1.0mg/mL时对大肠杆菌、白色假丝酵母和黑曲霉的抑菌圈直径分别为(21.84±0.28)、(16.33±0.54)、(11.54±0.51)mm。
Escherichia coli,Canidia albicans and Aspergillus niger were used as indicator strains for the bioassay-guided separation of antibacterial components from the ethyl acetate extract of fermentation broth of Paecilomyces militaris.The ethyl acetate extract was separated by silica gel column chromatography into two antibacterial fractions,Ⅴ and Ⅶ.Fraction Ⅴ was further separated by reversed-phase column chromatography to obtain a pure antibacterial compound,PMBA-1.At a concentration of 1.0 mg/mL,PMBA-1 presented an inhibition circle diameter of(16.21 ± 0.27) mm,(23.54 ± 0.35) mm and(27.33 ± 0.54) mm against Escherichia coli,Canidia albicans and Aspergillus niger,respectively.Another pure antibacterial compound,PMBA-2,was obtained from further separation of fraction Ⅶ by silica gel column chromatography.Its inhibition circle diameter at a concentration of 1.0 mg/mL was(21.84 ± 0.28),(16.33 ± 0.54) mm and(11.54 ± 0.51) mm for Escherichia coli,Canidia albicans and Aspergillus niger,respectively.
Escherichia coli,Canidia albicans and Aspergillus niger were used as indicator strains for the bioassay-guided separation of antibacterial components from the ethyl acetate extract of fermentation broth of Paecilomyces militaris.The ethyl acetate extract was separated by silica gel column chromatography into two antibacterial fractions,Ⅴ and Ⅶ.Fraction Ⅴ was further separated by reversed-phase column chromatography to obtain a pure antibacterial compound,PMBA-1.At a concentration of 1.0 mg/mL,PMBA-1 presented an inhibition circle diameter of(16.21 ± 0.27) mm,(23.54 ± 0.35) mm and(27.33 ± 0.54) mm against Escherichia coli,Canidia albicans and Aspergillus niger,respectively.Another pure antibacterial compound,PMBA-2,was obtained from further separation of fraction Ⅶ by silica gel column chromatography.Its inhibition circle diameter at a concentration of 1.0 mg/mL was(21.84 ± 0.28),(16.33 ± 0.54) mm and(11.54 ± 0.51) mm for Escherichia coli,Canidia albicans and Aspergillus niger,respectively.
引文
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[20]何亚琼,胡丰林,王利,等.一种被毛孢菌丝体中抗肿瘤活性组分的分离纯化和液质联用分析[J].菌物学报,2008,27(3):431-438.
[2]NAG T B,WANG H X.Pharmacological actions of Cordyceps,aprized folk medicine[J].Journal of Pharmacy and Phaumacology,2005,57:1509-1519.
[3]林群英,宋斌,李泰辉.蛹虫草研究进展[J].微生物学通报,2006,33(4):154-157.
[4]王建芳,杨春清.蛹虫草有效成分及药理作用研究进展[J].中医药信息,2005,22(5):30-32.
[5]WON S Y,PAR K E H.Anti-inflammatory and relate dpharmacological activities of cultured mucelia and fruiting bodies ofCordyceps militaris[J].Journal of Ethnopharmacology,2005,96(3):555-561.
[6]纪朋艳,罗速,崔新颖,等.中药蛹虫草的抗肿瘤活性及机制研究[J].北华大学学报,2005,6(4):324-329.
[7]LIN Y W,CHIANG B H.Anti-tumor activity of the fermentation brothof Cordyceps militaris cultured in the medium of Radix astragali[J].Process Biochemistry,2008,43(3):244-250.
[8]RAO Y K,FANG S H,WU W S,et al.Constituents isolated fromCordyceps militaris suppress enhanced inflammatory cediator’sproduction and human cancer cell proliferation[J].Journal ofEthnopharmacology,2010,131(2):363-367.
[9]JIANG Y,WONG J H,FU M,et al.Isolation of adenosine,isosinensetin and dimethylguanosine with antioxidant and HIV-1protease inhibiting activuties from fruiting bodies of Cordycepsmilitaris[J].Phytomedicine,2011,18(2):189-193.
[10]ZHANG Guoqing,HUANG Yuedong,BIAN Yong,et al.Hypoglycemic activity of the fungi Cordyceps militaris,Cordycepssinensis,Tricholoma mongolicum and Omphalia lapidescens instreptozotocin induced diabetic rats[J].Appl Microbiol Biotechnol,2006,72:1152-1156.
[11]姜泓,刘珂,孟舒,等.人工蛹虫草子实体化学成分[J].药学学报,2000,35(9):663-668.
[12]李维光,苏凤岩,黄荣年.北冬虫夏草菌丝体吨级液体深层发酵及生理活性物质的研究[J].微生物学杂志,1997,17(1):32-37.
[13]陈晋安,黄浩,郑忠辉,等.蛹虫草液体发酵条件的研究[J].集美大学学报,2001,6(3):219-223.
[14]MAO Xianbing,EKSRIWONG T,ZHONG Jianjiang,et al.Optimization of carbon source and carbon/nitrogen ratio for cordycepinproduction by submerged cultivation of medicinal mushroomCordyceps militaris[J].Process Biochemistry,2005,40:1667-1672.
[15]YEON S H,KIN J R,AHN Y J.Comparison of growth-inhibitingactivities of Cordyceps militaris and Paecilomyces japonica culturedon Bombyx mori pupae towards human gastrointestinal bacteria[J].Journal of the Science of Food and Agriculture,2007,87:54-59.
[16]王陶,邵颖,李文.蛹拟青霉不同菌株液体发酵代谢产物的抑菌活性研究[J].食品工业,2012(2):96-98.
[17]武忠伟,王运兵,赵现方,等.冬虫夏草和蛹虫草发酵液抗菌活性研究[J].微生物学杂志,2008,48(4):47-50.
[18]陈宏伟,朱蕴兰,邵颖,等.蛹虫草乙醇分级多糖生理功能研究[J].食品与发酵工业,2008,34(9):16-19.
[19]徐德峰,孙力军,王雅玲,等.蛹虫草抑菌活性研究开发现状与发展趋势[J].食品工业科技,2012,33(8):424-426.
[20]何亚琼,胡丰林,王利,等.一种被毛孢菌丝体中抗肿瘤活性组分的分离纯化和液质联用分析[J].菌物学报,2008,27(3):431-438.