Chlamydomonas reinhardtii-derived triple BmKbpp distorts membrane integrity for inhibiting bacterial growth

Abstract

Antibiotic abuses cause pathogen resistance to conventional antibiotics, threating public health globally. Antimicrobial peptides were considered as substitutes for antibiotic because of their broaden and rapid antibacterial activity. BmKbpp extracted from the Chinese scorpion Mesobuthus martensii Karsch inhibits the growth of gram-negative bacteria and fungi efficiently, while its efficacy for inhibiting the growth of the gram-positive Staphylococcus aureus was limited. Besides, how BmKbpp inhibits bacterial growth remains elusive thus far. In this study, a C-terminal hemagglutinin (HA) and 6×His (HA-6×His) tagged BmKbpp of three repeats (3×BmKbpp) was expressed in Chlamydomonas reinhardtii. The recombinant 3×BmKbpp-HA-6×His exhibited antibacterial activity against both gram-positive and gram-negative bacteria with a minimum inhibitory concentration between 20⁓200 µg/ml. The minimum bactericidal concentration ranged from 1 × MIC to 3 × MIC for 3×BmKbpp-HA-6×His. 3×BmKbpp-HA-6×His exhibited a good thermostability and resisted on pH shock and digestion of certain proteinases. It also displayed low cytotoxicity and hemolysis as tested on cell lines including HEK293, Vero, MDCK, and BHK-21. In addition, 3×BmKbpp-HA-6×His inhibits bacterial growth by effectively distorting bacterial cell membrane. Thus, C. reinhardtii-derived 3×BmKbpp-HA-6×His effectively inhibits bacterial growth, making it a promising antimicrobial peptide.