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从解淀粉芽孢杆菌 CBSYS12 中分离得到的一种新型抗菌肽 YS12 对耐药菌具有抗生物膜特性。

A novel antimicrobial peptide YS12 isolated from Bacillus velezensis CBSYS12 exerts anti-biofilm properties against drug-resistant bacteria.

机构信息

Department of Pharmacy, College of Pharmacy, Chosun University, Gwangju, 61452, Republic of Korea.

出版信息

Bioprocess Biosyst Eng. 2023 Jun;46(6):813-828. doi: 10.1007/s00449-023-02864-7. Epub 2023 Mar 30.

Abstract

Nowadays, the abuse of antibiotics has led to the rise of multi-drug-resistant bacteria. Antimicrobial peptides (AMPs), with broad-spectrum antimicrobial activity have attracted considerable attention as possible alternatives to traditional antibiotics. In this work, we aimed to evaluate the antimicrobial and anti-biofilm activity of an antimicrobial peptide designed as YS12 derived from Bacillus velezensis CBSYS12. The strain CBSYS12 was isolated from Korean food kimchi and purified followed by ultrafiltration and sequential chromatographic methodology. Hereafter, Tricine SDS-PAGE revealed a single protein band of around 3.3 kDa that was further confirmed in situ inhibitory activity of the gel. A similar molecular weight (~ 3348.4 Da) protein also appeared in MALDI-TOF confirming the purity and homogeneity of peptide YS12. Intriguingly, YS12 revealed a strong antimicrobial activity with a minimum inhibitory concentration (MIC) value ranging from 6 to 12 μg/ml for both Gram-positive and Gram-negative bacteria, such as E. coli, P. aeruginosa, MRSA 4-5, VRE 82, and M. smegmatis. We also determined the mode of action of the peptide against pathogenic microorganisms using different fluorescent dyes. In addition, the anti-biofilm assay demonstrated that peptide YS12 was able to inhibit biofilm formation  around 80% for both bacterial strains E. coli and P. aeruginosa at 80 µg/ml. Notably, YS12 exhibited a greater biofilm eradication activity than commercial antibiotics. In summary, our study proposed that peptide YS12 may be used as a promising therapeutic agent to overcome drug and biofilm-related infections.

摘要

如今,抗生素的滥用导致了多药耐药菌的出现。具有广谱抗菌活性的抗菌肽(AMPs)作为传统抗生素的替代品引起了相当大的关注。在这项工作中,我们旨在评估从韩国食品泡菜中分离出的芽孢杆菌 CBSYS12 中衍生的抗菌肽 YS12 的抗菌和抗生物膜活性。该菌株 CBSYS12 经超滤和连续层析方法分离和纯化。此后,Tricine SDS-PAGE 显示出约 3.3 kDa 的单一蛋白带,该蛋白带在凝胶原位抑制活性中得到进一步证实。MALDI-TOF 也显示出类似分子量(~3348.4 Da)的蛋白,证实了肽 YS12 的纯度和均一性。有趣的是,YS12 对革兰氏阳性和革兰氏阴性细菌(如大肠杆菌、铜绿假单胞菌、MRSA4-5、VRE82 和 M. smegmatis)均表现出很强的抗菌活性,最低抑菌浓度(MIC)值在 6 到 12 μg/ml 之间。我们还使用不同的荧光染料确定了该肽对病原微生物的作用模式。此外,抗生物膜试验表明,肽 YS12 能够在 80 μg/ml 时抑制大肠杆菌和铜绿假单胞菌两种菌株的生物膜形成约 80%。值得注意的是,YS12 表现出比商业抗生素更强的生物膜清除活性。总之,我们的研究表明,肽 YS12 可能被用作一种有前途的治疗剂,以克服与药物和生物膜相关的感染。

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