Recombinant expression of the precursor of rat lung surfactant protein B in Escherichia coli and its antibacterial mechanism

While the discovery of antibiotics has made a huge contribution to medicine, bacteria that are resistant to many antibiotics pose new challenges to medicine. Antimicrobial peptides (AMPs), a new kind of antibiotics, have attracted people's attention because they are not prone to drug resistance...

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Bibliographic Details
Published in:Protein expression and purification 2021-03, Vol.179, p.105801-105801, Article 105801
Main Authors: Cao, Wulong, Liu, Qin, Wang, Ting, Zhang, Qiuhan, Cheng, Fu, Tang, Yishan, Mei, Chenchen, Wen, Fang, Wang, Wanneng
Format: Article
Language:English
Subjects:
Antibacterial
Antibacterial peptide
Cytotoxicity
Mechanism
Pulmonary surfactant protein B precursor
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Summary:While the discovery of antibiotics has made a huge contribution to medicine, bacteria that are resistant to many antibiotics pose new challenges to medicine. Antimicrobial peptides (AMPs), a new kind of antibiotics, have attracted people's attention because they are not prone to drug resistance. In this study, glutathione transferase (GST) was used as a fusion partner to recombinantly expressed rat lung surfactant protein B precursor (proSP-B) in E. coli pLySs. Cck-8 evaluated the cytotoxicity of the fusion protein and calculated its 50% inhibitory concentration (IC50). The purified peptides showed broad-spectrum antibacterial activity using filter paper method and MIC, and propidium iodide (PI) was used to explore the antibacterial mechanism against Staphylococcus aureus. In addition, the pEGFP-N2-proSP-B vector was constructed to explore the localization of proSP-B in CCL-149 cells. We found that proSP-B has obvious antibacterial activity against Gram-positive bacteria, Gram-negative bacteria and fungi, and has broad-spectrum antibacterial activity. Besides, proSP-B fusion protein has low toxicity and can change the permeability of Staphylococcus aureus cell membrane to realize its antibacterial. For these reasons, proSP-B can be used as a potential natural antibacterial drug. •It was discovered that proSP-B was used as an antibacterial peptide and expressed by fusion with glutathione transferase.•The cytotoxicity of proSP-B fusion protein was tested for the first time.•The antibacterial spectrum and optimal pH of proSP-B fusion protein were determined for the first time.•ProSP-B can be located on the cell membrane of CCL-149 cells after being fused with green fluorescent protein (GFP).
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2020.105801