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The discovery and applications of antimicrobial peptides (AMPs) with antibacterial properties have been considered and proven as alternative antimicrobial agents to antibiotics. In this study, we have designed, produced, and purified a recombinant novel multifunctional hybrid antimicrobial peptide LL-37_Renalexin for the first time via the application of newly designed flexible GS peptide linker coupled with the use of our previously characterized small metal-binding proteins SmbP and CusF3H+ as carrier proteins that allow for an enhanced bacterial expression, using BL21(DE3) and SHuffle T7(DE3) <jats:italic>Escherichia coli<\/jats:italic> strains, and purification of the hybrid peptide via immobilized metal affinity chromatography. The purified tag-free LL-37_Renalexin hybrid peptide exhibited above 85% reduction in bacteria colony-forming units and broad-spectrum antimicrobial effects against <jats:italic>Staphylococcus aureus<\/jats:italic>, <jats:italic>Escherichia coli<\/jats:italic>, Methicillin-resistant <jats:italic>Staphylococcus aureus<\/jats:italic> (MRSA), and <jats:italic>Klebsiella pneumoniae<\/jats:italic> bacteria clinical isolates at a lower minimum inhibition concentration level (10\u201333 \u03bcM) as compared to its counterpart single-AMPs LL-37 and Renalexin (50\u2013100 \u03bcM).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Key points<\/jats:title>\n            <jats:p>\u2022 <jats:italic>The hybrid antimicrobial peptide LL-37_Renalexin has been designed using a GS linker.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\u2022 <jats:italic>The peptide was expressed with the carrier proteins SmbP and CusF3H+.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\u2022 <jats:italic>The hybrid peptide shows antibacterial potency against clinical bacterial isolates.<\/jats:italic>\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s00253-023-12887-5","type":"journal-article","created":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T09:02:51Z","timestamp":1705136571000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["LL-37_Renalexin hybrid peptide exhibits antimicrobial activity at lower MICs than its counterpart single peptides"],"prefix":"10.1007","volume":"108","author":[{"given":"Julius Kwesi","family":"Narh","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nestor G.","family":"Casillas-Vega","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9824-3961","authenticated-orcid":false,"given":"Xristo","family":"Zarate","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,1,13]]},"reference":[{"key":"12887_CR1","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1007\/s10529-019-02656-8","volume":"41","author":"J Adam\u00edkov\u00e1","year":"2019","unstructured":"Adam\u00edkov\u00e1 J, Anto\u0161ov\u00e1 M, Polakovi\u010d M (2019) Chromatographic purification of recombinant human erythropoietin. 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