贝加尔唐松草内生真菌的分离纯化及其次级代谢产物生物活性研究
发布时间：2019-06-10 07:38
【摘要】：近年来药用植物内生真菌是得到新的活性天然产物及研发新药的潜在资源,具有重要的研究和开发利用价值。甘肃省药用植物资源丰富、分布广泛,但关于药用植物内生真菌的研究报道较少,因此,本论文以甘肃省贝加尔唐松草作为研究材料,从其各组织分离纯化得到内生真菌,初步研究内生真菌次级代谢产物的抗氧化、抗菌活性,研究内容和结果如下:采用组织分离法,从药用植物贝加尔唐松草(Thalictrum baicalense Turcz.)的根,茎,叶,种子中共分离得到18株内生真菌,根据形态进行初步鉴定,分属于曲霉属、顶孢霉属、镰刀菌属、香柱菌属、链格孢菌属、匍柄霉属,其中镰刀菌属为优势属,占33.33%。制备18株内生真菌的次级代谢产物,利用加热回流法对干燥菌丝体进行提取,提取液减压蒸干得菌丝体乙醇提取部位;用乙酸乙酯和正丁醇分别对发酵液进行萃取,各萃取3次,合并每次萃取液,减压蒸干,得到发酵液乙酸乙酯和正丁醇萃取部位。采用DPPH自由基清除法、总还原力法、羟基自由基清除法和微量肉汤稀释法对贝加尔唐松草内生真菌发酵产物进行抗氧化及体外抑菌活性研究。抗氧化结果表明,菌株发酵液的乙酸乙酯部位普遍具有抗氧化活性,其中菌株Y-3对DPPH自由基清除活性最强,达到91.04%,其活性仅次于Vc,菌株J-2对羟基自由基清除活性最大,达到77.86%,菌株Z-6总还原力较好,Vc的相当含量为190.2μg/mg。抑菌活性结果表明,菌株Z-1乙酸乙酯部位抑菌活性最好,对金黄色葡萄球菌和地衣芽孢杆菌的MIC值为125μg/mL,对肺炎克雷伯菌、大肠杆菌、无乳链球菌、铜绿假单胞菌的MIC值均为250μg/mL;菌株Z-3和Z-4正丁醇部位抑菌活性较好,对6种供试菌的MIC值都达到250μg/mL;菌株Z-7乙醇部位抑菌活性较好,对金黄色葡萄球菌、肺炎克雷伯菌、大肠杆菌和铜绿假单胞菌的MIC值均为500μg/mL,对无乳链球菌和地衣芽孢杆菌MIC值为1000μg/m L。利用硅胶和Sephadex LH-20凝胶柱层析,并结合薄层色谱检识,从唐松草内生真菌J-2发酵液的乙酸乙酯部位分离纯化得到化合物,通过单体化合物的理化性质以及核磁共振氢谱、核磁共振碳谱、质谱等现代波谱学手段对其结构进行了鉴定,初步鉴定为1-O-十六碳烷酰-2-O-(9Z-十八碳烯酰)-3-O-(9Z,12Z-十八碳二烯酰)甘油酯(1),其余化合物的的鉴定仍在继续,由于时间关系剩余部位的分离、化合物结构鉴定和单体化合物生物活性测定将在后续工作中进行。
[Abstract]:In recent years, endophytic fungi in medicinal plants are potential resources for obtaining new active natural products and developing new drugs, which have important research, development and utilization value. Gansu Province is rich in medicinal plant resources and widely distributed, but there are few reports on endophytic fungi of medicinal plants. Therefore, in this paper, the endophytic fungi were isolated and purified from the tissues of Pinus tabulaeformis in Gansu Province. The antioxidant and antibacterial activities of secondary metabolites of endophytic fungi were studied. The contents and results were as follows: the medicinal plant (Thalictrum baicalense Turcz.) was isolated by tissue isolation method. A total of 18 endophytic fungi were isolated from roots, stems, leaves and seeds. According to their morphology, they belonged to Aspergillus, Fusarium, columella, Streptomyces and stolymyces, among which Fusarium was the dominant genus. It accounted for 33.33%. The secondary metabolites of 18 strains of endophytic fungi were prepared. The dried fungi were extracted by heating and reflux method, and the ethanol extraction parts of the fungi were obtained by vacuum evaporation of the extract. Ethyl acetate and n-butanol were used to extract the fermentation broth for three times, and the extraction solution was combined with each extraction solution and evaporated under pressure to obtain the extraction parts of ethyl acetate and n-butanol in the fermentation broth. DPPH radical scavenging method, total reduction method, hydroxyl radical scavenging method and micro broth dilution method were used to study the antioxidant and in vitro bacteriostatic activities of endophytic fungal fermentation products of Pinus tabulaeformis. The results of antioxidant activity showed that the ethyl acetate part of the fermentation broth of the strain generally had antioxidant activity, among which strain Y 鈮，
本文编号：2496290
[Abstract]:In recent years, endophytic fungi in medicinal plants are potential resources for obtaining new active natural products and developing new drugs, which have important research, development and utilization value. Gansu Province is rich in medicinal plant resources and widely distributed, but there are few reports on endophytic fungi of medicinal plants. Therefore, in this paper, the endophytic fungi were isolated and purified from the tissues of Pinus tabulaeformis in Gansu Province. The antioxidant and antibacterial activities of secondary metabolites of endophytic fungi were studied. The contents and results were as follows: the medicinal plant (Thalictrum baicalense Turcz.) was isolated by tissue isolation method. A total of 18 endophytic fungi were isolated from roots, stems, leaves and seeds. According to their morphology, they belonged to Aspergillus, Fusarium, columella, Streptomyces and stolymyces, among which Fusarium was the dominant genus. It accounted for 33.33%. The secondary metabolites of 18 strains of endophytic fungi were prepared. The dried fungi were extracted by heating and reflux method, and the ethanol extraction parts of the fungi were obtained by vacuum evaporation of the extract. Ethyl acetate and n-butanol were used to extract the fermentation broth for three times, and the extraction solution was combined with each extraction solution and evaporated under pressure to obtain the extraction parts of ethyl acetate and n-butanol in the fermentation broth. DPPH radical scavenging method, total reduction method, hydroxyl radical scavenging method and micro broth dilution method were used to study the antioxidant and in vitro bacteriostatic activities of endophytic fungal fermentation products of Pinus tabulaeformis. The results of antioxidant activity showed that the ethyl acetate part of the fermentation broth of the strain generally had antioxidant activity, among which strain Y 鈮，
本文编号：2496290
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