一株黄芪内生真菌甾醇类代谢产物的分离和鉴定
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摘要
目的:研究分离自夏季黄芪根部的内生真菌AR-17-S03的次级代谢产物。方法:采用组织学的方法从恒山黄芪中分离内生真菌,真菌的鉴定采用形态学法结合分子生物学法,采用硅胶柱色谱法、薄层层析制备色谱(pTLC)、Sephadex LH-20凝胶柱色谱法和半制备HPLC色谱法分离内生真菌代谢产物,采用质谱法、核磁共振法结合物化性质鉴定分离得到的9个化合物。结果:经形态学结合分子生物学鉴定该菌株黑曲霉,为9个甾醇类化合物鉴定为麦角甾醇、过氧化麦角甾醇、胆固醇、啤酒甾醇、23-甲基麦角甾-7,22-二烯-3β,5α,9α-三醇、5α,8α-表二氧麦角甾-6,9(11),22-三烯-3β-醇、5α,8α-表二氧-23-甲基麦角甾-6,22-二烯-3β-醇、麦角甾-4,6,8(14),22-四烯-3-酮、3β,5α-二羟基麦角甾-7,22-二烯-6-酮。结论:其中,23-甲基麦角甾-7,22-二烯-3β,5α,9α-三醇、5α,8α-表二氧-23-甲基麦角甾-6,22-二烯-3β-醇、麦角甾-4,6,8(14),22-四烯-3-酮是首次从黄芪内生真菌中分离得到。
Objective:Study on secondary metabolites from endophytic fungus AR-17-S03 isolated from the roots of Astragalus membranaceus in summer. Methods:Endophyic fungus was isolated from Astragalus membranaceus by the method of histology. The fungus was identified by morphological method combined with molecular biology method. The compounds were isolated by column chromatography,pTLC,Sephadex LH-20 and pre-HPLC. And they were identified by the spectral analysis of mass spectrometry and nuclear magnetic resonance. Results:Endophytic fungi AR-17-S03 isolated from roots of Astragalus membranaceus were identified as Aspergillus niger. Nine sterols were obtained and identified as ergosterol,ergosterol peroxide,cholesterol,cerevistero,23-methylergosta-7,22-diene-3β,5α,9α-triol,5α,8α-epidioxy-ergosta-6,9(11),22-trien-3β-ol,5α,8α-epidioxy-23-methylergosta-6,22-diene-3β-ol,ergosta-4,6,8(14),22-tetraen-3-one and 3β,5α-dihydroxy-ergosta-7,22-diene-6-one. Conclusion:23-methylergosta-7,22-diene-3β,5α,9α-triol,5α,8α-epidioxy-23-methylergosta-6,22-diene-3β-ol and ergosta-4,6,8(14),22-tetraen-3-one were obtained from the endophytic fungus of Astragalus membranaceus for the first time.
Objective:Study on secondary metabolites from endophytic fungus AR-17-S03 isolated from the roots of Astragalus membranaceus in summer. Methods:Endophyic fungus was isolated from Astragalus membranaceus by the method of histology. The fungus was identified by morphological method combined with molecular biology method. The compounds were isolated by column chromatography,pTLC,Sephadex LH-20 and pre-HPLC. And they were identified by the spectral analysis of mass spectrometry and nuclear magnetic resonance. Results:Endophytic fungi AR-17-S03 isolated from roots of Astragalus membranaceus were identified as Aspergillus niger. Nine sterols were obtained and identified as ergosterol,ergosterol peroxide,cholesterol,cerevistero,23-methylergosta-7,22-diene-3β,5α,9α-triol,5α,8α-epidioxy-ergosta-6,9(11),22-trien-3β-ol,5α,8α-epidioxy-23-methylergosta-6,22-diene-3β-ol,ergosta-4,6,8(14),22-tetraen-3-one and 3β,5α-dihydroxy-ergosta-7,22-diene-6-one. Conclusion:23-methylergosta-7,22-diene-3β,5α,9α-triol,5α,8α-epidioxy-23-methylergosta-6,22-diene-3β-ol and ergosta-4,6,8(14),22-tetraen-3-one were obtained from the endophytic fungus of Astragalus membranaceus for the first time.
引文
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[18]Yaoita Y,Endo M,Tani Y,et al. Sterol constituents from seven mushrooms[J]. Chemical & Pharmaceutical Bulletin,1999,47:847-851.
[19]Luo X,Li F M,Shinde P B,et al. 26,27-cyclosterols and other polyoxygenated sterols from a marine sponge Topsentia sp[J]. Journal of Natural Products,2006,69:1760-1768.
[20]Yasunori Y,Keiko A,Hiroyuki O,et al. Sterol constituents from five edible mushrooms[J]. Chemical & Pharmaceutical Bulletin,1998,46:944-950.
[21]Gaspar E M M,Neves H J C. Steroidal constituents from mature wheat straw[J]. Phytochemistry,1993,34:523-527.
[2]Schaller H. Review:The role of sterols in plant growth and development[J]. Progress in Lipid Research,2003,42(3):163-175.
[3]金俊,卢梦瑶,厉秋岳,等. 以甾醇为底物微生物法合成甾体类化合物的研究进展[J]. 食品研究与开发,2018,39(10):205-209.
[4]Zhou F,Zhang H C,Liu R,et al. Isolation and biological evaluation of secondary metabolites of the endophyticfungus Aspergillus fumigatus from Astragalus membranaceus[J]. Chemistry of Natural Compounds,2013,49:568-570.
[5]张弘弛,刘瑞,李慧,等. 拮抗皮革霉变的黄芪内生真菌AR07 抑菌成分的分离鉴定[J]. 中国皮革,2017,46(5):32-36.
[6]刘瑞,张弘弛,李慧,等. 一株恒山产蒙古黄芪内生真菌Penicillium griseofulvum次生代谢产物的分离和菌种鉴定[J]. 中国实验方剂学杂志,2016,22(16):25-29.
[7]张弘弛,刘瑞,李慧,等. 黄芪内生真菌中甲醛降解菌的筛选及降解特性[J]. 西北农业学报,2017,26(2):311-316.
[8]张弘弛,刘瑞,李慧,等. 黄芪内生真菌AR15中吲哚二酮哌嗪及其对黄芪白粉病原菌的抑制作用[J]. 西北林学院学报,2017,32(1):197-202.
[9]王维,马养民,张弘弛,等. 黑果枸杞内生真菌E21菌株次生代谢产物的研究[J]. 中国新药杂志,2013,22(4):460-464.
[10]梁小蕊,苗凤萍,宋银平,等. 海藻内生真菌棘孢木霉dl-34中的二萜和甾体化合物[J]. 化学与生物工程,2016,33(5):32-36.
[11]陶美华,颜健,魏孝义,等. 白木香内生真菌Fimetariella rabenhorstii的甾体类代谢产物[J]. 热带亚热带植物学报,2011,19(1):75-78.
[12]丁唯嘉,陈洁怡,王锦华,等. 2株红树内生真菌共培养产生的甾醇类化合物的分离鉴定[J]. 华南农业大学学报,2013,34(2):267-271.
[13]杨瑞云,李春远,林永成,等. 南海马尾藻(Sargassum sp.)内生真菌No.ZZF36中甾醇化合物的研究[J]. 中药材,2006,29(9):908-909.
[14]郦皆秀,李进,徐丽珍,等. 西藏产冬虫夏草化学成分研究[J]. 中国药学杂志,2003(38):499-501.
[15]Leslie G A A,Yalamanchili G,Francis J S,et al. Minor and trace sterols in marine invertebrates.26.isolation and structure elucidation of nine new 5α,8α-epidioxy sterols from four marine organisms[J]. Journal of Organic Chemistry,1981,46:3860-3866.
[16]孟艳辉,苏镜娱,曾陇梅. 海绵Hircinia variabilis化学成分研究[J]. 中山大学学报:自然科学版,1993,31:90-92.
[17]朱峰,刘晓红,温露,等. 南海海洋真菌1924号代谢产物的研究[J]. 中山大学学报:自然科学版,2005,44:66-69.
[18]Yaoita Y,Endo M,Tani Y,et al. Sterol constituents from seven mushrooms[J]. Chemical & Pharmaceutical Bulletin,1999,47:847-851.
[19]Luo X,Li F M,Shinde P B,et al. 26,27-cyclosterols and other polyoxygenated sterols from a marine sponge Topsentia sp[J]. Journal of Natural Products,2006,69:1760-1768.
[20]Yasunori Y,Keiko A,Hiroyuki O,et al. Sterol constituents from five edible mushrooms[J]. Chemical & Pharmaceutical Bulletin,1998,46:944-950.
[21]Gaspar E M M,Neves H J C. Steroidal constituents from mature wheat straw[J]. Phytochemistry,1993,34:523-527.