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菌株PBR11,一种具有抗菌潜力的森林衍生土壤。

sp. Strain PBR11, a Forest-Derived Soil with Antimicrobial Potential.

作者信息

Mazumdar Rajkumari, Dutta Partha Pratim, Saikia Juri, Borah Jagat Chandra, Thakur Debajit

机构信息

Life Sciences Division, Institute of Advanced Study in Science and Technology, Guwahati, India.

Department of Molecular Biology and Biotechnology, Cotton University, Guwahati, India.

出版信息

Microbiol Spectr. 2023 Jan 31;11(2):e0348922. doi: 10.1128/spectrum.03489-22.

Abstract

The isolate PBR11 was isolated from the forest rhizosphere soil of Pobitora Wildlife Sanctuary (PWS), Assam, India. The isolate was identified as sp. with 92.91% sequence similarity to their closest type strain, Streptomyces atrovirens NRRL B-16357 DQ026672. The strain demonstrated significant antimicrobial activity against 19 test pathogens, including multidrug-resistant (MDR) clinical isolates and dermatophytes. Phenol, 2,5-bis(1,1-dimethylethyl), is the major chemical compound detected by gas chromatography-mass spectrometry in the ethyl acetate extract of PBR11 (EtAc-PBR11). The presence of the PKS type II gene (type II polyketide synthases) and chitinase gene suggested that it has been involved in the production of antimicrobial compounds. Metabolic profiling of the EtAc-PBR11 was performed by thin-layer chromatography and flash chromatography resulted in the extraction of two bioactive fractions, namely, PBR11Fr-1 and PBR11Fr-2. Liquid chromatography-tandem mass spectrometry analysis of both the fractions demonstrated the presence of significant antimicrobial compounds, including ethambutol. This is the first report on the detection of antituberculosis drug in the bioactive fractions of sp. PBR11. EtAc-PBR11 and PBR11Fr-1 showed the lowest MIC values (>0.097 and >0.048 μg/mL, respectively) against Candida albicans MTCC 227, whereas they showed the highest MIC values (>0.390 and >0.195 μg/mL, respectively) against Escherichia coli ATCC BAA-2469. The effects of PBR11Fr-1 were investigated on the pathogens by using a scanning electron microscope. The results indicated major morphological alterations in the cytoplasmic membrane. PBR11Fr-1 exhibited low cytotoxicity on normal hepatocyte cell line (CC-1) and the percent cell viability started to decline as the concentration increased from 50 μg/mL (87.07% ± 3.22%) to 100 μg/mL (81.26% ± 2.99%). Novel antibiotic breakthroughs are urgently required to combat antimicrobial resistance. are the principal producers of antibiotics. The present study demonstrated the broad-spectrum antimicrobial potential of an strain sp. strain PBR11 isolated from the PWS of Assam, India, which represents diverse, poorly screened habitats for novel microorganisms. The strain displayed 92.4% sequence similarity with genes of the closest type strain, indicating that the strain may represent a novel taxon within the phylum . The metabolomics studies of EtAc-PBR11 revealed structurally diverse antimicrobial agents, including the detection of the antituberculosis drug ethambutol, in the bioactive fraction of sp. PBR11 for the first time. The PBR11 strain also yielded positive results for the antibiotic synthesis gene and the chitinase gene, both of which are responsible for broad-spectrum antimicrobial activity. This suggests that the untouched forest ecosystems have a tremendous potential to harbor potent actinomycetia for future drug discovery.

摘要

分离株PBR11是从印度阿萨姆邦波比托拉野生动物保护区(PWS)的森林根际土壤中分离出来的。该分离株被鉴定为链霉菌属,与最接近的模式菌株深绿链霉菌NRRL B - 16357 DQ026672的序列相似性为92.91%。该菌株对19种测试病原体表现出显著的抗菌活性,包括多重耐药(MDR)临床分离株和皮肤癣菌。2,5 - 双(1,1 - 二甲基乙基)苯酚是气相色谱 - 质谱法在PBR11乙酸乙酯提取物(EtAc - PBR11)中检测到的主要化合物。II型聚酮合酶(PKS)基因和几丁质酶基因的存在表明它参与了抗菌化合物的产生。通过薄层色谱对EtAc - PBR11进行代谢谱分析,快速色谱法提取出两个生物活性组分,即PBR11Fr - 1和PBR11Fr - 2。对这两个组分进行液相色谱 - 串联质谱分析,结果表明存在包括乙胺丁醇在内的显著抗菌化合物。这是首次在链霉菌属PBR11的生物活性组分中检测到抗结核药物的报道。EtAc - PBR11和PBR11Fr - 1对白色念珠菌MTCC 227显示出最低的最低抑菌浓度值(分别>0.097和>0.048μg/mL),而对大肠杆菌ATCC BAA - 2469显示出最高的最低抑菌浓度值(分别>0.390和>0.195μg/mL)。使用扫描电子显微镜研究了PBR11Fr - 1对病原体的影响。结果表明细胞质膜出现了主要的形态变化。PBR11Fr - 1对正常肝细胞系(CC - 1)表现出低细胞毒性,随着浓度从50μg/mL(87.07%±3.22%)增加到100μg/mL(81.26%±2.99%),细胞活力百分比开始下降。迫切需要新的抗生素突破来对抗抗菌耐药性。放线菌是抗生素的主要生产者。本研究证明了从印度阿萨姆邦PWS分离出的链霉菌属菌株PBR11具有广谱抗菌潜力,该地区代表了多样且筛选较少的新型微生物栖息地。该菌株与最接近的模式菌株的基因序列相似性为92.4%,表明该菌株可能代表放线菌门内的一个新分类单元。对EtAc - PBR11的代谢组学研究首次揭示了链霉菌属PBR11生物活性组分中结构多样的抗菌剂,包括抗结核药物乙胺丁醇的检测。PBR11菌株的抗生素合成基因和几丁质酶基因也呈阳性结果,这两个基因都负责广谱抗菌活性。这表明未受干扰的森林生态系统具有蕴藏强大放线菌用于未来药物发现的巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da0b/10101066/0c020ee172ed/spectrum.03489-22-f001.jpg

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