Exogenous Production of N-acetylmuramyl-L Alanine Amidase (LysM2) from Siphoviridae Phage Affecting Anti-Gram-Negative Bacteria: Evaluation of Its Structure and Function

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 -hosted Siphoviridae phage (SPP1 phage, Subtilis Phage Pavia 1) was exogenously expressed in . The sequences of genes encoding peptidoglycan hydrolase...

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Bibliographic Details
Published in:Avicenna journal of medical biotechnology 2022-01, Vol.14 (1), p.46-53
Main Authors: Miri, Morteza, Yazdianpour, Sepideh, Abolmaali, Shamsozoha, Darvish Alipour Astaneh, Shakiba
Format: Article
Language:English
Subjects:
Amino acids
Antibacterial agents
Enzymes
Escherichia coli
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Summary: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 -hosted Siphoviridae phage (SPP1 phage, Subtilis Phage Pavia 1) was exogenously expressed in . The sequences of 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 . Furthermore, the structure and function of LysM2 was evaluated . The optimized gene was synthesized, subcloned in the pET28a, and expressed in BL21(DE3). The antibacterial effects of the protein on the peptidoglycan substrates were studied. , on 816 gene encoding a 33 protein was confirmed as specific SPP1 phage enzyme. The enzyme is composed of 271 amino acids, with a half-life of 10 in . 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 (sp) by turbidity, and an antibacterial activity against , and in agar diffusion assays. Taken together, our results confirmed that LysM2 is an inhibiting agent for gram-negative bacteria.
ISSN:2008-2835
2008-4625
DOI:10.18502/ajmb.v14i1.8169