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评价产自尼泊尔喜马拉雅红豆杉内生真菌的生物活性化合物的抗菌、抗氧化和细胞毒性特性。

Evaluation of antimicrobial, antioxidant and cytotoxic properties of bioactive compounds produced from endophytic fungi of Himalayan yew ( ) in Nepal.

机构信息

Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel, Kavrepalanchok District, Bagmati, 45200, Nepal.

Department of Forest Genetics and Plant Physiology, Umeå Plant Science Centre, Swedish University of Agricultural Sciences, Umeå, SE-901 83, Sweden.

出版信息

F1000Res. 2020 May 19;9:379. doi: 10.12688/f1000research.23250.2. eCollection 2020.

Abstract

Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study, which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties. The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts. Out of 16 different trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera: and . The ethyl acetate extracts isolated from three endophytic fungi: , and showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of and also showed promising antimicrobial activity against , and with a minimum inhibitory concentration of 250μg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC of 104.2 and 125.9µg/ml, respectively. The crude fungal extracts obtained from endophytes: , and upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.

摘要

内生真菌在天然生物活性产物的发现中很大程度上尚未得到充分探索,尽管它们是具有潜在药用前景的新型化合物的丰富来源。本研究对具有巨大药用价值的内生真菌多样性及其产生生物活性次生代谢产物的能力进行了研究,通过分析抗氧化、抗菌和细胞毒性特性来实现。使用基因组 DNA 样本通过 ITS-PCR 鉴定内生真菌。使用乙酸乙酯溶剂萃取法提取次生代谢产物。通过薄层层析法、总酚含量(TPC)、总黄酮含量(TFC)和 DPPH 测定法分析抗氧化活性,通过琼脂孔扩散法分析抗菌活性。用卤虫致死试验分析真菌提取物的细胞毒性。从多帕特安不同地点采集的 16 种不同树木中分离出 13 种不同的内生真菌,并分为 9 个不同属: 、 、 、 、 、 、 、 、 。从三种内生真菌: 、 、 中分离出的乙酸乙酯提取物表现出显著的 TPC 值,分别为 204±6.144、312.3±2.147 和 152.7±4.958µg GAE/mg 干提取物,TFC 值分别为 177.9±2.911、644.1±4.202 和 96.38±3.851µg RE/mg 干提取物。此外,这三种提取物均表现出与浓度呈依赖性的自由基清除活性,IC 浓度分别为 22.85、22.15 和 23.001µg/ml。提取物 和 也表现出对 、 和 的良好抗菌活性,所有细菌的最低抑菌浓度均为 250μg/ml。两种样品对虾幼体均表现出细胞毒性,LC 分别为 104.2 和 125.9µg/ml。从内生真菌: 、 、 中获得的粗真菌提取物经过纯化和进一步鉴定生物活性化合物后,可能成为新型药物制剂的迷人来源。

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