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热嗜甲基菌噬菌体 MMP17 来源的 MMPphg 作为一种潜在的抗革兰氏阴性和革兰氏阳性细菌的抗菌剂。

MMPphg from the thermophilic Meiothermus bacteriophage MMP17 as a potential antimicrobial agent against both Gram-negative and Gram-positive bacteria.

机构信息

Faculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming, 650500, Yunnan Province, China.

Engineering Research Center for Replacement Technology of Feed Antibiotics of Yunnan College, 727 South Jingming Road, Kunming, 650500, Yunnan Province, China.

出版信息

Virol J. 2020 Aug 25;17(1):130. doi: 10.1186/s12985-020-01403-0.

Abstract

BACKGROUND

New strategies are urgently needed to deal with the growing problem of multidrug-resistant bacterial pathogens. As the natural viruses against bacteria, recently, bacteriophages have received particular attention. Here, we identified and characterized a novel peptidoglycan hydrolase named MMPphg by decoding the complete genome sequence of Meiothermus bacteriophage MMP17, which was isolated in Tengchong hot spring in China and contains a circular genome of 33,172 bp in size and a GC content of 63.4%.

FINDINGS

We cloned the MMPphg gene, overproduced and purified the phage lytic protein, which contains a highly conserved M23 metallopeptidase domain and can be activated by Mg and Zn. MMPphg is capable of withstanding temperatures up to 70 °C, and preserved more than 80% of its activity after a 30 min treatment between 35 and 65 °C. More interestingly, by disrupting bacterial cells, MMPphg exhibits surprising antimicrobial activity against both Gram-negative and Gram-positive pathogenic bacteria, especially antibiotic-resistant strains such as Escherichia coli O157, Staphylococcus aureus and Klebsiella pneumonia.

CONCLUSIONS

In the current age of mounting antibiotic resistance, these results suggest the great potential of MMPphg, the gene product of bacteriophage MMP17, in combating bacterial infections and shed light on bacteriophage-based strategies to develop alternatives to conventional antibiotics for human or veterinary applications.

摘要

背景

急需新策略来应对日益严重的多药耐药细菌病原体问题。作为天然的细菌病毒,噬菌体最近受到了特别关注。在这里,我们通过解码中国腾冲温泉中分离的梅奥thermus 噬菌体 MMP17 的完整基因组序列,鉴定并表征了一种新型的肽聚糖水解酶,命名为 MMPphg。该噬菌体含有一个大小为 33172bp 的圆形基因组,GC 含量为 63.4%。

结果

我们克隆了 MMPphg 基因,大量生产并纯化了噬菌体裂解蛋白,该蛋白含有一个高度保守的 M23 金属蛋白酶结构域,可以被 Mg 和 Zn 激活。MMPphg 能耐受高达 70°C 的温度,在 35 至 65°C 之间处理 30 分钟后,仍能保持超过 80%的活性。更有趣的是,通过破坏细菌细胞,MMPphg 对革兰氏阴性和革兰氏阳性的致病性细菌,特别是大肠杆菌 O157、金黄色葡萄球菌和肺炎克雷伯菌等抗生素耐药菌株,表现出惊人的抗菌活性。

结论

在当前抗生素耐药性日益严重的情况下,这些结果表明噬菌体 MMP17 基因产物 MMPphg 在对抗细菌感染方面具有巨大的潜力,并为基于噬菌体的策略提供了启示,以开发替代传统抗生素的方法,用于人类或兽医应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2908/7448439/f8719e3e87d8/12985_2020_1403_Fig1_HTML.jpg

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