摘要
【目的】通过化学诱导方法从一株海洋真菌Hypocrea lixii中获得多样化的活性小分子化合物。【方法】对菌株在盐酸普鲁卡因的化学诱导作用下进行固体发酵,代谢产物采用硅胶减压柱层析、Sephadex LH-20以及pHPLC等手段分离纯化,对化合物运用NMR、MS等波谱学技术并比对文献进行结构鉴定,采用Ellman比色法及DPPH自由基清除法对化合物进行初步抗乙酰胆碱酯酶(AChE)和抗氧化活性测试。【结果】从海洋真菌Hypocrea lixii在盐酸普鲁卡因诱导下的发酵提取物中分离得到4个异黄酮类化合物,包括6''-O-crotonylgenistin(1)、Genistein(2)、Daidzein(3)和Genistin(4),以及对氨基苯甲酸甲酯(5)、对羟基苯丙酸(6)和胞嘧啶(7),其中化合物1和6为该种中首次报道,化合物5为生物转化来源首次报道,化合物2~4可能源自培养基。在100μmol/L剂量浓度下,化合物1~4对AChE的抑制以及DPPH自由基的清除活性均较弱,化合物5和6显示了相对强的AChE抑制活性,抑制率分别为40%和38%。【结论】盐酸普鲁卡因对该菌次级代谢产生了显著的影响,为深入研究该菌株化学诱导次级代谢产物提供基础。
【Objective】This study aimed at discovering diverse bioactive small molecular compounds from a marine fungus Hypocrea lixii by chemical induction.【Methods】The marine fungus Hypocrea lixii was fermented in solid culture medium induced by procaine hydrochloride.The metabolites were purified by vacuum liquid chromatography,Sephadex LH-20 and pHPLC.The structures were elucidated by the analysis of spectroscopic data and comparison to literatures.All compounds were tested for acetylcholinesterase(AChE)inhibitory and antioxidant activity through Ellman and DPPH free radical scavenging assay.【Results】Seven compounds were isolated from the procaine-induced metabolites of Hypocrea lixii including four isoflavones(6''-O-crotonylgenistin,genistein,daidzein and genistin),methyl 4-aminobenzoate,phloretic acid,and cytosine.For the first time,compounds 1 and 6 are reported from this species,compound 5 is reported as biotransformation product,and compounds 2~4 may come from the medium.In bioassay with the dose concentration of 100μmol/L,compounds 1~4 only displayed weak AChE inhibitory and antioxidant activities;whereas compounds 5 and 6 showed relatively stronger AChE inhibitory activity with inhibition ratios of 40%and 38%,respectively.【Conclusion】Procaine hydrochloride exerted significant influence on the secondary metabolism of this strain.The present study lays the foundation for further study on its chemically induced secondary metabolites.
作者
杨文聪
刘亚月
杨静明
黎钊坪
梁金月
聂影影
马小翔
张翼
YANG Wen-cong;LIU Ya-yue;YANG Jing-ming;LI Zhao-ping;LIANG Jin-yue;NIE Ying-ying;MA Xiao-xiang;ZHANG Yi(Shenzhen Insititute of Guangdong Ocean University,Shenzhen 518120,China;College of Food Science and Technology,Guangdong Ocean University,Key Laboratory of Guangdong Province Aquatic Products Processing and Safety,Guangdong Province Engineering Laboratory for Marine Biological Products,Guangdong Provincial Engineering Technology Research Center of Marine Food,Key Laboratory of Advanced Processing of Aquatic Products of Guangdong Higher Education Institution,Institute for Marine Drugs and Nutrition,Guangdong Ocean University,Zhanjiang 524088,China)
出处
《广东海洋大学学报》
CAS
2020年第1期55-63,共9页
Journal of Guangdong Ocean University
基金
广东省扬帆计划引进紧缺拔尖人才项目(201433009)
国家自然科学基金(21807015)
广东省自然科学基金(2018A030307046)
深圳市科创委基础研究自由探索项目(JCYJ20170306165013264)
广东省应用科技研发重大专项(2016B020235001)
广东省促进经济发展专项资金(海洋经济发展用途)项目(粤自然资源合[2019] 015号)
深圳市大鹏新区产业发展资金(KY20180203及PT201901-05)
湛江市海洋渔业局项目(A18018)
