{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["14(1)"],"submitter":["Miri M"],"pubmed_abstract":["<h4>Background</h4>To obtain endolysin with impact(s) on gram-negative bacteria as well as gram-positive bacteria, N-acetylmuramyl L-alanine-amidase (MurNAc-LAA) from a <i>Bacillus subtilis</i>-hosted Siphoviridae phage (SPP1 phage, Subtilis Phage Pavia 1) was exogenously expressed in <i>Escherichia coli (E. coli)</i>.<h4>Methods</h4>The sequences of <i>MurNAc-LAA</i> genes encoding peptidoglycan hydrolases were obtained from the Virus-Host database. The sequence of MurNAc-LAA was optimized by GenScript software to generate MurNAc-LAA-MMI (LysM2) for optimal expression in <i>E. coli</i>. Furthermore, the structure and function of LysM2 was evaluated <i>in silico</i>. The optimized gene was synthesized, subcloned in the pET28a, and expressed in <i>E. coli</i> BL21(DE3). The antibacterial effects of the protein on the peptidoglycan substrates were studied.<h4>Results</h4><i>LysM2</i>, on 816 <i>bp</i> gene encoding a 33 <i>kDa</i> protein was confirmed as specific SPP1 phage enzyme. The enzyme is composed of 271 amino acids, with a half-life of 10 <i>hr</i> in <i>E. coli</i>. <i>In silico</i> analyses showed 34.2% alpha-helix in the secondary structure, hydrophobic N-terminal, and lysine-rich C-terminal, and no antigenic properties in LysM2 protein. This optimized endolysin revealed impacts against <i>Proteus</i> (sp) by turbidity, and an antibacterial activity against <i>Klebsiella pneumoniae, Salmonella typhimurium</i>, and <i>Proteus vulgaris</i> in agar diffusion assays.<h4>Conclusion</h4>Taken together, our results confirmed that LysM2 is an inhibiting agent for gram-negative bacteria."],"journal":["Avicenna journal of medical biotechnology"],"pagination":["46-53"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9017466"],"repository":["biostudies-literature"],"pubmed_title":["Exogenous Production of N-acetylmuramyl-L Alanine Amidase (LysM2) from Siphoviridae Phage Affecting Anti-Gram-Negative Bacteria: Evaluation of Its Structure and Function."],"pmcid":["PMC9017466"],"pubmed_authors":["Miri M","Darvish Alipour Astaneh S","Yazdianpour S","Abolmaali S"],"additional_accession":[]},"is_claimable":false,"name":"Exogenous Production of N-acetylmuramyl-L Alanine Amidase (LysM2) from Siphoviridae Phage Affecting Anti-Gram-Negative Bacteria: Evaluation of Its Structure and Function.","description":"<h4>Background</h4>To obtain endolysin with impact(s) on gram-negative bacteria as well as gram-positive bacteria, N-acetylmuramyl L-alanine-amidase (MurNAc-LAA) from a <i>Bacillus subtilis</i>-hosted Siphoviridae phage (SPP1 phage, Subtilis Phage Pavia 1) was exogenously expressed in <i>Escherichia coli (E. coli)</i>.<h4>Methods</h4>The sequences of <i>MurNAc-LAA</i> genes encoding peptidoglycan hydrolases were obtained from the Virus-Host database. The sequence of MurNAc-LAA was optimized by GenScript software to generate MurNAc-LAA-MMI (LysM2) for optimal expression in <i>E. coli</i>. Furthermore, the structure and function of LysM2 was evaluated <i>in silico</i>. The optimized gene was synthesized, subcloned in the pET28a, and expressed in <i>E. coli</i> BL21(DE3). The antibacterial effects of the protein on the peptidoglycan substrates were studied.<h4>Results</h4><i>LysM2</i>, on 816 <i>bp</i> gene encoding a 33 <i>kDa</i> protein was confirmed as specific SPP1 phage enzyme. The enzyme is composed of 271 amino acids, with a half-life of 10 <i>hr</i> in <i>E. coli</i>. <i>In silico</i> analyses showed 34.2% alpha-helix in the secondary structure, hydrophobic N-terminal, and lysine-rich C-terminal, and no antigenic properties in LysM2 protein. This optimized endolysin revealed impacts against <i>Proteus</i> (sp) by turbidity, and an antibacterial activity against <i>Klebsiella pneumoniae, Salmonella typhimurium</i>, and <i>Proteus vulgaris</i> in agar diffusion assays.<h4>Conclusion</h4>Taken together, our results confirmed that LysM2 is an inhibiting agent for gram-negative bacteria.","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Jan-Mar","modification":"2025-04-18T12:11:18.232Z","creation":"2025-04-06T21:49:23.936Z"},"accession":"S-EPMC9017466","cross_references":{"pubmed":["35509364"],"doi":["10.18502/ajmb.v14i1.8169"]}}