Abstract
Staphylococcus aureus is a Gram-positive pathogen that is capable of infecting almost every organ in the human body. Alarmingly, the rapid emergence of methicillin-resistant S. aureus strains (MRSA) jeopardizes the available treatment options. Herein, we propose sustainable, low-cost production of recombinant lysostaphin (rLST), which is a native bacteriocin destroying the staphylococcal cell wall through its endopeptidase activity. We combined the use of E. coli BL21(DE3)/pET15b, factorial design, and simple Ni-NTA affinity chromatography to optimize rLST production. The enzyme yield was up to 50 mg/L culture, surpassing reported systems. Our rLST demonstrated superlative biofilm combating ability by inhibiting staphylococcal biofilms formation and detachment of already formed biofilms, compared to vancomycin and linezolid. Furthermore, we aimed at developing a novel rLST topical formula targeting staphylococcal skin infections. The phase inversion composition (PIC) method fulfilled this aim with its simple preparatory steps and affordable components. LST nano-emulgel (LNEG) was able to extend active LST release up to 8 h and cure skin infections in a murine skin model. We are introducing a rapid, convenient rLST production platform with an outcome of pure, active rLST incorporated into an effective LNEG formula with scaling-up potential to satisfy the needs of both research and therapeutic purposes.
Highlights
Since their clinical introduction in the 1930s, antibiotics have highly impacted human morbidity and mortality
Staphylococcal strains used in this study included: S. aureus strains Newman [16] and USA300, which is a multiple antibiotic resistant and community-acquired strain [17], the S. simulans strain TNK3 [18], the S. epidermidis strain ATCC 12228 (ATCC), and the S. lugdunensis strain N920143 [19]
Luria Bertani (LB) was supplemented with ampicillin at a final concentration of 100 μg/mL
Summary
Since their clinical introduction in the 1930s, antibiotics have highly impacted human morbidity and mortality. Decades of antibiotics mis-use has applied selective pressure on pathogens, which results in unprecedented antimicrobial resistance [1,2]. S. aureus is a leading cause of hospital and community infections worldwide [3]. The most recent antimicrobial for combating staphylococcal infections was discovered 30 years ago. To fill this gap in a way that considerably lower the resistance potential, new non-conventional ways had to be explored. Investigated alternatives include plant-derived compounds [4], bacteriophages and phage lysins [5], RNA-based therapeutics [6], antimicrobial adjuvants [7], and antimicrobial peptides (AMP) [8]
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