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利用宏基因组纳米孔测序结合液相色谱-质谱分析探索土壤分离株MZ921932产生的抗菌代谢产物的性质。

Exploring the Nature of the Antimicrobial Metabolites Produced by Soil Isolate MZ921932 Using a Metagenomic Nanopore Sequencing Coupled with LC-Mass Analysis.

作者信息

Eltokhy Mohamed A, Saad Bishoy T, Eltayeb Wafaa N, Yahia Ibrahim S, Aboshanab Khaled M, Ashour Mohamed S E

机构信息

Department of Microbiology, Faculty of Pharmacy, Misr International University (MIU), Cairo 19648, Egypt.

Department of Bioinformatics, HITS Solutions Co., Cairo 11765, Egypt.

出版信息

Antibiotics (Basel). 2021 Dec 22;11(1):12. doi: 10.3390/antibiotics11010012.

Abstract

The continuous emergence of multidrug-resistant (MDR) pathogens poses a global threat to public health. Accordingly, global efforts are continuously conducted to find new approaches to infection control by rapidly discovering antibiotics, particularly those that retain activities against MDR pathogens. In this study, metagenomic nanopore sequence analysis coupled with spectroscopic methods has been conducted for rapid exploring of the various active metabolites produced by soil isolate. Preliminary soil screening resulted in selection of a Gram-positive isolate identified via 16S ribosomal RNA gene sequencing as MZ921932. The isolate showed a broad range of activity against MDR Gram-positive, Gram-negative, and spp. A metagenomics sequence analysis of the soil sample harboring isolate MZ921932 (NCBI GenBank accession PRJNA785410) revealed the presence of conserved biosynthetic gene clusters of petrobactin, tridecaptin, locillomycin (β-lactone), polymyxin, and macrobrevin (polyketides). The liquid chromatography/mass (LC/MS) analysis of the metabolites confirmed the presence of petrobactin, locillomycin, and macrobrevin. In conclusion, isolate MZ921932 is a promising rich source for broad spectrum antimicrobial metabolites. The metagenomic nanopore sequence analysis was a rapid, easy, and efficient method for the preliminary detection of the nature of the expected active metabolites. LC/MS spectral analysis was employed for further confirmation of the nature of the respective active metabolites.

摘要

多重耐药(MDR)病原体的不断出现对全球公共卫生构成了威胁。因此,全球一直在努力通过快速发现抗生素,特别是那些对MDR病原体仍具有活性的抗生素,来寻找新的感染控制方法。在本研究中,已进行了宏基因组纳米孔序列分析并结合光谱方法,以快速探索土壤分离株产生的各种活性代谢产物。初步土壤筛选结果选定了一株革兰氏阳性分离株,通过16S核糖体RNA基因测序鉴定为MZ921932。该分离株对MDR革兰氏阳性菌、革兰氏阴性菌和 spp.表现出广泛的活性。对含有分离株MZ921932的土壤样本(NCBI GenBank登录号PRJNA785410)进行的宏基因组序列分析显示,存在petrobactin、tridecaptin、locillomycin(β-内酯)、多粘菌素和macrobrevin(聚酮化合物)的保守生物合成基因簇。对 代谢产物的液相色谱/质谱(LC/MS)分析证实了petrobactin、locillomycin和macrobrevin的存在。总之,分离株MZ921932是广谱抗菌代谢产物的一个有前景的丰富来源。宏基因组纳米孔序列分析是一种快速简便且高效的方法,用于初步检测预期活性代谢产物的性质。LC/MS光谱分析用于进一步确认各个活性代谢产物的性质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23c3/8773065/01e884e0673a/antibiotics-11-00012-g001.jpg

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