Plectasin: from evolution to truncation, expression, and better druggability

Xuan Li; Ya Hao; Na Yang; Ruoyu Mao; Da Teng; Jianhua Wang

doi:10.3389/fmicb.2023.1304825

Plectasin: from evolution to truncation, expression, and better druggability

Front Microbiol. 2023 Dec 21:14:1304825. doi: 10.3389/fmicb.2023.1304825. eCollection 2023.

Authors

Xuan Li^{1

2

3}, Ya Hao^{1

2

3}, Na Yang^{1

2

3}, Ruoyu Mao^{1

2

3}, Da Teng^{1

2

3}, Jianhua Wang^{1

2

3}

Affiliations

¹ Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
² Innovative Team of Antimicrobial Peptides and Alternatives to Antibiotics, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
³ Key Laboratory of Feed Biotechnology, Chinese Academy of Agricultural Sciences, Department of Agriculture and Rural Affairs, Beijing, China.

Abstract

Non-computational classical evolution analysis of plectasin and its functional relatives can especially contribute tool value during access to meet requirements for their better druggability in clinical use. Staphylococcus aureus is a zoonotic pathogen that can infect the skin, blood, and other tissues of humans and animals. The impact of pathogens on humans is exacerbated by the crisis of drug resistance caused by the misuse of antibiotics. In this study, we analyzed the evolution of anti-Staphylococcus target functional sequences, designed a series of plectasin derivatives by truncation, and recombinantly expressed them in Pichia pastoris X-33, from which the best recombinant Ple-AB was selected for the druggability study. The amount of total protein reached 2.9 g/L following 120 h of high-density expression in a 5-L fermenter. Ple-AB was found to have good bactericidal activity against gram-positive bacteria, with minimum inhibitory concentration (MIC) values ranging between 2 and 16 μg/mL. It showed good stability and maintained its bactericidal activity during high temperatures, strong acid and alkali environments. Notably, Ple-AB exhibited better druggability, including excellent trypsin resistance, and still possessed approximately 50% of its initial activity following exposure to simulated intestinal fluids for 1 h. In vitro safety testing of Ple-AB revealed low hemolytic activity against mouse erythrocytes and cytotoxicity against murine-derived macrophages. This study successfully realized the high expression of a new antimicrobial peptide (AMP), Ple-AB, in P. pastoris and the establishment of its oral administration as an additive form with high trypsin resistance; the study also revealed its antibacterial properties, indicating that truncation design is a valuable tool for improving druggability and that the candidate Ple-AB may be a novel promising antimicrobial agent.

Keywords: druggability; evolution; expression; plectasin; truncation.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the National Natural Science Foundation of China (Grant Nos. 31702146 and 31872393), the AMP Direction of the National Innovation Program of Agricultural Science and Technology in CAAS (Grant CAAS-ASTIP-2013-FRI-02), and the Key Project of Alternatives to Antibiotics for Feed (Grant CAAS-ZDXT2018008) and Animal (Grant CAAS-ZDRW202111) Usages—the National Agricultural Science and Technology Innovation Program.