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化学诱导对一株海洋来源土曲霉C23-3次生代谢产物及其生物活性的影响
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广东省扬帆计划引进紧缺拔尖人才项目(201433009);广东省自然科学基金(2018A030307046);广东海洋大学科研启动项目(E15155);深圳市科创委基础研究自由探索项目(JCYJ20170306165013264);食品微生物学课程群广东省省级优秀教学团队(粤教高函[2015]133号);广东海洋大学优秀教学团队(校人事[2017]18号)


Influence of chemical induction on the secondary metabolites and biological activities of a marine-derived fungal strain Aspergillus terreus C23-3
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    摘要:

    【背景】海洋微生物是药理活性先导化合物的重要源泉,而化学诱导是深入挖掘其次生代谢潜力的一种便捷手段。【目的】以一株海洋来源土曲霉C23-3为出发菌株,筛选出可促进其产生更丰富代谢产物(包括丁内酯类化合物)的发酵条件,进一步开发菌株在抗阿尔茨海默症方面的潜力。【方法】分别选用海水马铃薯培养基、麦芽浸膏培养基、大米培养基和大豆培养基4种基础培养基,利用丁酸钠、辛二酰苯胺异羟肟酸(Suberoylanilide hydroxamic acid,SAHA)、5-氮杂胞嘧啶核苷(5-azacytidine,5-azaC)、盐酸普鲁卡因(Procaine hydrochloride)、ZnCl2和CuCl2共6种化学诱导剂对土曲霉C23-3进行微量的诱导培养。观察该菌菌丝体形态变化,并根据薄层层析(Thin-layer chromatography,TLC)指纹图谱、化学显色及生物活性自显影结果初筛出代谢产物丰富且乙酰胆碱酯酶抑制活性与抗氧化活性产物丰富的诱导条件,并进行进一步常量发酵。采用上述手段及高效液相色谱-二极管阵列检测器(High performance liquid chromatography-Diode array detector,HPLC-DAD)分析常量发酵代谢产物的总体多样性及丁内酯类化合物的多样性。【结果】发现CuCl2、ZnCl2、SAHA和5-azaC可引起土曲霉C23-3菌丝体生长形态显著变化,海水马铃薯培养基+100 μmol/L丁酸钠等6种诱导条件在微量培养初筛及常量发酵复筛中均可使该菌株产生多样化的活性代谢产物,其丁内酯类代谢产物也具有一定差异性。【结论】化学诱导手段可促使土曲霉C23-3产生丰富且新颖的活性代谢产物,为多样化的抗阿尔茨海默症天然产物的发现奠定了基础。

    Abstract:

    [Background] Marine microorganisms are important source of pharmaceutical active leading compounds and chemical induction can be used as a convenient approach to mine their secondary metabolic potential. [Objective] Based on a marine-derived fungus Aspergillus terreus C23-3, this study aimed to optimize fermentation conditions for more abundant metabolites (including butyrolactones), and exploit the potential of the strains in drugs targeting Alzheimer’s disease. [Methods] The strain was first cultivated on micro-scale, by taking four basic cultures mediums including seawater potato medium, malt extract medium, rice medium and soybean medium, and adopting six chemical inducers including sodium butyrate, suberoylanilide hydroxamic acid (SAHA), 5-azacytidine (5-azaC), procaine hydrochloride, ZnCl2 and CuCl2. The morphological variation of the mycelia was observed. Then, based on the thin layer chromatography (TLC) fingerprints, chemical coloration and bioautography results, inducing conditions with rich new metabolites and rich anti-acetylcholinesterase and antioxidant products were initially screened out for further routine-scale fermentation. Using the above methods and high-performance liquid chromatography-diode array detector (HPLC-DAD) analysis, the diversity of the total metabolites and of butyrolactones was analyzed for the routine fermentation. [Results] The results indicated that CuCl2, ZnCl2, SAHA and 5-azaC caused remarkable changes in the growth morphology of Aspergillus terreus C23-3 mycelium. Six chemical inducing conditions, e.g. seawater potato medium plus 100 μmol/L sodium butyrate, led to diverse bioactive metabolites both in micro-scale cultivation and routine-scale fermentation. Furthermore, its butyrolactone metabolites also displayed difference under different conditions. [Conclusion] Chemical induction approaches can prompt the Aspergillus terreus C23-3 to produce rich bioactive substances, laying foundation for the discovery of diverse anti-Alzheimer’s disease natural products.

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杨静明,杨文聪,刘亚月,聂影影,雷晓凌,张翼. 化学诱导对一株海洋来源土曲霉C23-3次生代谢产物及其生物活性的影响[J]. 微生物学通报, 2019, 46(3): 441-452

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  • 在线发布日期: 2019-03-01
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