biostudies-literatureVarotsou CHellenic Foundation for Research and Innovation8523https://www.ebi.ac.uk/biostudies/studies/S-EPMC10218239biostudies-literatureUnknown24(10)The emergence of multidrug-resistant (MDR) bacteria has risen rapidly, leading to a great threat to global public health. A promising solution to this problem is the exploitation of phage endolysins. In the present study, a putative N-acetylmuramoyl-L-alanine type-2 amidase (NALAA-2, EC 3.5.1.28) from <i>Propionibacterium</i> bacteriophage PAC1 was characterized. The enzyme (<i>Pa</i>Ami1) was cloned into a T7 expression vector and expressed in <i>E. coli</i> BL21 cells. Kinetics analysis using turbidity reduction assays allowed the determination of the optimal conditions for lytic activity against a range of Gram-positive and negative human pathogens. The peptidoglycan degradation activity of <i>Pa</i>Ami1 was confirmed using isolated peptidoglycan from <i>P. acnes</i>. The antibacterial International journal of molecular sciencesCharacterization and Engineering Studies of a New Endolysin from the <i>Propionibacterium acnes</i> Bacteriophage PAC1 for the Development of a Broad-Spectrum Artilysin with Altered Specificity.PMC102182394036Labrou NEPremetis GEVarotsou CfalseCharacterization and Engineering Studies of a New Endolysin from the <i>Propionibacterium acnes</i> Bacteriophage PAC1 for the Development of a Broad-Spectrum Artilysin with Altered Specificity.The emergence of multidrug-resistant (MDR) bacteria has risen rapidly, leading to a great threat to global public health. A promising solution to this problem is the exploitation of phage endolysins. In the present study, a putative N-acetylmuramoyl-L-alanine type-2 amidase (NALAA-2, EC 3.5.1.28) from <i>Propionibacterium</i> bacteriophage PAC1 was characterized. The enzyme (<i>Pa</i>Ami1) was cloned into a T7 expression vector and expressed in <i>E. coli</i> BL21 cells. Kinetics analysis using turbidity reduction assays allowed the determination of the optimal conditions for lytic activity against a range of Gram-positive and negative human pathogens. The peptidoglycan degradation activity of <i>Pa</i>Ami1 was confirmed using isolated peptidoglycan from <i>P. acnes</i>. The antibacterial 2023-01-01T00:00:00Z2023 May2026-04-07T17:35:56.055Z2024-11-21T02:00:38.813ZS-EPMC102182393723987410.3390/ijms24108523