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短梗霉 F3,一种新型的来源于南方红豆杉果实的紫杉醇产生内生真菌:分离、鉴定、紫杉醇产量提高和抗肿瘤活性。

Alternaria alternata F3, a Novel Taxol-Producing Endophytic Fungus Isolated from the Fruits of Taxus cuspidata: Isolation, Characterization, Taxol Yield Improvement, and Antitumor Activity.

机构信息

Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry University, Harbin, 150040, People's Republic of China.

Engineering Research Center of Forest Bio-Preparation, Ministry of Education, Northeast Forestry University, Harbin, 150040, People's Republic of China.

出版信息

Appl Biochem Biotechnol. 2024 Apr;196(4):2246-2269. doi: 10.1007/s12010-023-04661-0. Epub 2023 Jul 27.

Abstract

In this study, a novel taxol-producing endophytic fungus, strain F3, was isolated from the fruits of Taxus cuspidata and identified as Alternaria alternata according to its macroscopic and microscopic traits and sequence analysis of internal transcribed spacer (ITS). The presence of taxol was detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) and confirmed by ultra-high-performance liquid chromatography-electrospray coupled to tandem mass spectrometry (UPLC-ESI-MS/MS) and nuclear magnetic resonance (NMR). The fermentation parameters of strain F3 were then optimized for high taxol production. The maximum taxol yield of 195.4 µg L by A. alternata F3 was observed in 200-mL yeast peptone dextrose (YPD) broth, at an initial pH value of 6.0, supplemented with 0.1 g L sodium acetate, 0.25 g L salicylic acid, and 0.00125 g L silver nitrate and inoculum size 2%, and incubated at 28 °C and 150 rpm for 8 days, which was 2.12-fold compared with the initial yield of taxol. Also, fungal taxol exhibited antitumor activity towards human lung carcinoma (A549) cell line and human cervical carcinoma (Hela) cell line with IC values of 3.98 µg mL and 0.35 µg mL. Overall, this is the first report on taxol-producing endophytic fungus isolated from the fruits of Taxus. This study offers a novel source for the production of taxol for anticancer treatment.

摘要

在这项研究中,从红豆杉果实中分离出一株新型紫杉醇产生内生真菌 F3,根据其宏观和微观特征以及内部转录间隔区(ITS)序列分析,鉴定为交替单胞菌。通过薄层层析(TLC)和高效液相色谱(HPLC)检测到紫杉醇的存在,并通过超高效液相色谱-电喷雾串联质谱(UPLC-ESI-MS/MS)和核磁共振(NMR)进行了确认。然后优化了菌株 F3 的发酵参数以实现高产紫杉醇。在初始 pH 值为 6.0、添加 0.1 g L 乙酸钠、0.25 g L 水杨酸和 0.00125 g L 硝酸银以及接种量为 2%的 200-mL 酵母胰蛋白胨葡萄糖(YPD)培养基中,A. alternata F3 的最大紫杉醇产量为 195.4 μg L,在 28°C 和 150 rpm 下培养 8 天,与紫杉醇的初始产量相比提高了 2.12 倍。此外,真菌紫杉醇对人肺癌(A549)细胞系和人宫颈癌(Hela)细胞系表现出抗肿瘤活性,IC 值分别为 3.98 μg mL 和 0.35 μg mL。总的来说,这是首次从红豆杉果实中分离出产紫杉醇的内生真菌。本研究为紫杉醇的生产提供了一种新的抗癌治疗来源。

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