Kumar Pradeep, Singh Balwant, Thakur Vikram, Thakur Abhishek, Thakur Nandita, Pandey Deepak, Chand Duni
Department of Biotechnology, Himachal Pradesh University, Shimla, Himachal Pradesh, India.
Faculty of Applied Sciences and Biotechnology, Shoolini University, Solan, Himachal Pradesh, India.
Biotechnol Rep (Amst). 2019 Nov 15;24:e00395. doi: 10.1016/j.btre.2019.e00395. eCollection 2019 Dec.
Taxol® (generic name Paclitaxel) is a chemotherapeutic drug, effective against head, neck, breast, lung, bladder, ovary, and cervix cancers. Rising demands in chemotherapy and limited supply of natural taxol have ultimately increased the cost of the drug. Semi synthesis using taxol precursors is not able to meet the global supply and has intensified the need to find alternative ways of taxol production. In the present study, 34 different endophytes were isolated from sp. collected from Shimla, Himachal Pradesh (India). Primary screening of taxol-producing fungi was carried out based on the presence of gene, essential for the taxol biosynthetic pathway. A fungal isolate TPF-06 was screened to be a taxol-producing strain based on the PCR amplification results. It was characterized and identified as by (Accession No. KU-837249). Multiple sequence alignment (MSA) of nuclear ribosomal internal transcribed spacer (ITS) region and phylogenetic analysis confirmed that strain belonged to clade (Accession No. MF-374798) and is endophytic in nature. Presence of taxol was detected and quantified by High-Performance Liquid Chromatography (HPLC) and characterized by using Thin Layer Chromatography (TLC), Ultraviolet (UV) spectroscopy, Mass spectrometry (MS), Fourier-Transform Infrared Spectroscopy (FTIR) and Nuclear Magnetic Resonance (NMR) spectroscopy. Microbial fermentation in the S7 medium yielded 1.60 g/L of taxol, which to the best of our knowledge is the highest taxol production from an endophytic fungus. Findings of the present study suggest that the is an excellent alternate source for taxol supply, and it may become a highly potent strain on a commercial scale. The involvement of gene in KU-837249 strain further suggested a way of increasing taxol yield in fungi by medium engineering and recombinant DNA technology in the future.
紫杉醇(通用名:帕罗西汀)是一种化疗药物,对头部、颈部、乳腺、肺部、膀胱、卵巢和宫颈癌有效。化疗需求的增加以及天然紫杉醇供应的有限最终推高了该药物的成本。使用紫杉醇前体的半合成方法无法满足全球供应,因此迫切需要寻找生产紫杉醇的替代方法。在本研究中,从印度喜马偕尔邦西姆拉采集的 植物中分离出 34 种不同的内生菌。基于紫杉醇生物合成途径中必需的 基因的存在,对产紫杉醇真菌进行了初步筛选。根据 PCR 扩增结果,筛选出一株真菌分离株 TPF - 06 为产紫杉醇菌株。通过 鉴定为 (登录号:KU - 837249)。核糖体内部转录间隔区(ITS)区域的多序列比对(MSA)和系统发育分析证实该菌株属于 进化枝(登录号:MF - 374798),且本质上是内生菌。通过高效液相色谱(HPLC)检测和定量紫杉醇的存在,并使用薄层色谱(TLC)、紫外(UV)光谱、质谱(MS)、傅里叶变换红外光谱(FTIR)和核磁共振(NMR)光谱对其进行表征。在 S7 培养基中进行微生物发酵产生了 1.60 g/L 的紫杉醇,据我们所知,这是内生真菌产生的最高紫杉醇产量。本研究结果表明, 是紫杉醇供应的极佳替代来源,并且在商业规模上可能成为一种高效菌株。KU - 837249 菌株中 基因的参与进一步提示了未来通过培养基工程和重组 DNA 技术提高真菌中紫杉醇产量的途径。