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宏基因组学方法在从红树林沉积物中发现和开发新型生物活性化合物8-去甲氧基-10-脱氧斯特夫霉素中的应用

Metagenomics approaches in the discovery and development of new bioactive compound of 8-demethoxy-10-deoxysteffimycin from mangrove sediments.

作者信息

Sujith P, Singh J Rajesh, Jayalakshmi S, Kandaswamy Karthikeyan, Shaik Mohammed Rafi, Hussain Shaik Althaf, Kari Zulhisyam Abdul, Guru Ajay

机构信息

Department of Microbiology, SRM Arts and Science College, Kattankulathur, Chengalpattu District, Tamil Nadu 603203 India.

C.A.S in Marine Biology, Annamalai University, Parangipettai, Tamil Nadu 608002 India.

出版信息

3 Biotech. 2024 Dec;14(12):303. doi: 10.1007/s13205-024-04137-w. Epub 2024 Nov 17.

DOI:10.1007/s13205-024-04137-w
PMID:39564541
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11570573/
Abstract

A metagenomic library consisting of 15,000 clones was constructed from the mangrove sediment. An antimicrobially active clone from the metagenomic library PS49 was identified by function- based screening. This paper presents the results of the biochemical characterization and metagenomic library screening of the marine-derived antibiotic, 8-demethoxy-10-deoxysteffimycin. Plasmid libraries were constructed, and clones were produced using a metagenomic approach. Out of 15,000 clones, 81 clones were screened for antimicrobial activity, and five potential clones were selected. The activity of one clone was characterized and named as PS49. The bioactive compounds from the selected clone were checked for antimicrobial, antioxidant, and anticancer activities. The clone PS49 was tested against various pathogens including bacteria and fungi and it showed inhibitory effects against all the tested pathogens. The antimicrobially active fractions were then crystallized and subjected to spectroscopic analysis such as FTIR, NMR and LC-MS analysis. The substance from clone PS49 has finally been recognized, and the compound from clone PS49 has been identified as 8-demethoxy-10-deoxysteffimycin. The substances isolated from the PS49 clone exhibited strong anticancer activity against skin cancer-cell lines SK-MEL2. The compounds showed a reduction in cell viability with an increase in the compound concentration. The compounds obtained from clone PS49 showed an IC value of 85 µg/ml.

摘要

从红树林沉积物中构建了一个由15000个克隆组成的宏基因组文库。通过基于功能的筛选,从宏基因组文库PS49中鉴定出一个具有抗菌活性的克隆。本文介绍了海洋来源抗生素8-去甲氧基-10-脱氧司替霉素的生化特性及宏基因组文库筛选结果。构建了质粒文库,并采用宏基因组方法产生克隆。在15000个克隆中,筛选了81个克隆的抗菌活性,选择了5个潜在克隆。对其中一个克隆的活性进行了表征,并命名为PS49。检测了所选克隆的生物活性化合物的抗菌活性、抗氧化活性和抗癌活性。对克隆PS49针对包括细菌和真菌在内的各种病原体进行了测试,结果表明它对所有测试病原体均有抑制作用。然后将具有抗菌活性的组分结晶,并进行FTIR、NMR和LC-MS分析等光谱分析。最终识别出了来自克隆PS49的物质,克隆PS49中的化合物已被鉴定为8-去甲氧基-10-脱氧司替霉素。从PS49克隆中分离出的物质对皮肤癌细胞系SK-MEL2表现出很强的抗癌活性。随着化合物浓度的增加,细胞活力降低。从克隆PS49获得的化合物的IC值为85μg/ml。

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