Gp29 LysA of mycobacteriophage TM4 can hydrolyze peptidoglycan through an N-acetyl-muramoyl-L-alanine amidase activity

Urdániz, Estefanía; Martin, Mariano; Payaslián, Florencia; Defelipe, Lucas A.; Dodes, Martín; Martínez, Mariano; Alzari, P.M.; Cabrera, Gabriela M.; Martí, Marcelo A.; Piuri, Mariana

doi:10.1016/j.bbapap.2021.140745

Cited by 4 publications

(4 citation statements)

References 48 publications

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“…At room temperature, lysozyme can rapidly hydrolyze the substrate as soon as five minutes, while our proteins take at least two hours to hydrolyze the PG. This is consistent with studies in endolysins from mycobacteriophage (amidase) and enterobacterial phage (lysozyme) which were reported to take 1-2h to degrade PG [135,136]. Note that the hydrolysis assay performed in this study was carried out under the same conditions (PBS pH 7.4, 37°C) for all proteins.…”

Section: Pg-degrading Activitiessupporting

confidence: 90%

Investigating Novel Streptomyces Bacteriophage Endolysins as Potential Antimicrobial Agents

Maneekul

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As antibiotic resistance has become a major global threat, the World Health Organization has urgently called scientists for alternative strategies for control of bacterial infections. Endolysin, a protein encoded by a phage gene, can degrade bacterial peptidoglycan (PG). Currently, there are three endolysin products in the clinical phase. We, thus, are interested in exploring novel endolysins from Streptomyces phages as only a few of them have been experimentally characterized. Using bioinformatics tools, we identified nine functional domain groups from 250 Streptomyces phages putative endolysins. NootNoot gp34 (transglycosylase; Nt34lys), Nabi gp26 (amidase; Nb26lys), Tribute gp42 (PGRP; Tb42lys), and LazerLemon gp35 (CHAP; LL35lys) were selected for experimental studies. We hypothesized that (1) the proteins of interest will have the ability to degrade PG, and (2) the proteins will be potential antimicrobial agents against ESKAPE safe relatives. The results showed that LL35lys, Nb26lys and Tb42lys exhibit PG-degrading activity on zymography and hydrolysis assay. The enzymes (400 µg/mL) can reduce PG turbidity to 32-40%. The killing assay suggested that Tb42lys possess a boarder range (Escherichia coli, Pseudomonas putida, Acinetobacter baylyi and Klebsiella aerogenes). While Nb26lys can attack Gram-negative bacteria, LL35lys can only reduce the growth of the Gram-positive strains with an MIC90 of 2 µg/mL. A higher concentration (≥300 µg/mL) of Nb26lys is needed to treat P. putida and K. aerogenes. Therefore, endolysins from Streptomyces phage have potential as possible antimicrobial agents against ESKAPE bacteria.

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Section: Pg-degrading Activitiessupporting

confidence: 90%

Investigating Novel Streptomyces Bacteriophage Endolysins as Potential Antimicrobial Agents

Maneekul

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“…However, their low stability can hinder their clinical use. Endolysins from mesophilic bacteria infecting phages generally lose their hydrolytic activity above 45–55 °C (Premetis et al 2023a ; Abouhmad et al 2020 ; Urdániz et al 2022 ). Researchers have attempted to fuse thermophile lytic enzymes with CWBDs to create thermostable endolysins (Swift et al 2019 ).…”

Section: Discussionmentioning

confidence: 99%

“…Alternative methods include structure-based point mutations, in silico design of chimeric endolysins, and stabilizer addition to improve thermostability (Heselpoth et al 2015 ; Love et al 2021 ). The melting temperature ( T m ) of Ami1 indicates high thermal stability, compared with other known bacteriophage endolysins (Linden et al 2015 ; Bustamante et al 2012 ; Abouhmad et al 2020 ; Urdániz et al 2022 ), suggesting that Ami1 can be classified into thermostable enzymes.…”

Section: Discussionmentioning

confidence: 99%

Metagenomic analysis of hot spring soil for mining a novel thermostable enzybiotic

Pantiora,

Georgakis,

Premetis

et al. 2024

Appl Microbiol Biotechnol

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The misuse and overuse of antibiotics have contributed to a rapid emergence of antibiotic-resistant bacterial pathogens. This global health threat underlines the urgent need for innovative and novel antimicrobials. Endolysins derived from bacteriophages or prophages constitute promising new antimicrobials (so-called enzybiotics), exhibiting the ability to break down bacterial peptidoglycan (PG). In the present work, metagenomic analysis of soil samples, collected from thermal springs, allowed the identification of a prophage-derived endolysin that belongs to the N-acetylmuramoyl-L-alanine amidase type 2 (NALAA-2) family and possesses a LysM (lysin motif) region as a cell wall binding domain (CWBD). The enzyme (Ami1) was cloned and expressed in Escherichia coli, and its bactericidal and lytic activity was characterized. The results indicate that Ami1 exhibits strong bactericidal and antimicrobial activity against a broad range of bacterial pathogens, as well as against isolated peptidoglycan (PG). Among the examined bacterial pathogens, Ami1 showed highest bactericidal activity against Staphylococcus aureus sand Staphylococcus epidermidis cells. Thermostability analysis revealed a melting temperature of 64.2 ± 0.6 °C. Overall, these findings support the potential that Ami1, as a broad spectrum antimicrobial agent, could be further assessed as enzybiotic for the effective treatment of bacterial infections. Key points • Metagenomic analysis allowed the identification of a novel prophage endolysin • The endolysin belongs to type 2 amidase family with lysin motif region • The endolysin displays high thermostability and broad bactericidal spectrum

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“…Enzymes that destroy peptidoglycan are also secreted by bacteria in the competition be-tween bacteria of different lines for the development of new habitat niches and are strictly species-specific [15][16][17]. Bacteriophages also use murein hydrolases and lysozyme-like proteins to penetrate bacteria, destroying their peptidoglycan [18][19][20]. Lysozyme-like proteins that degrade peptidoglycan to disaccharide-containing derivatives have also been found in fungi [21].…”

Section: Sources Of Muramyl Peptides In the Human's Bodymentioning

confidence: 99%

Regulation of Immune Homeostasis via Muramyl Peptides-Low Molecular Weight Bioregulators of Bacterial Origin

Guryanova

2022

Microorganisms

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Metabolites and fragments of bacterial cells play an important role in the formation of immune homeostasis. Formed in the course of evolution, symbiotic relationships between microorganisms and a macroorganism are manifested, in particular, in the regulation of numerous physiological functions of the human body by the innate immunity receptors. Low molecular weight bioregulators of bacterial origin have recently attracted more and more attention as drugs in the prevention and composition of complex therapy for a wide range of diseases of bacterial and viral etiology. Signaling networks show cascades of causal relationships of deterministic phenomena that support the homeostasis of multicellular organisms at different levels. To create networks, data from numerous biomedical and clinical research databases were used to prepare expert systems for use in pharmacological and biomedical research with an emphasis on muramyl dipeptides. Muramyl peptides are the fragments of the cell wall of Gram-positive and Gram-negative bacteria. Binding of muramyl peptides with intracellular NOD2 receptors is crucial for an immune response on pathogens. Depending on the microenvironment and duration of action, muramyl peptides possess positive or negative regulation of inflammation. Other factors, such as genetic, pollutions, method of application and stress also contribute and should be taken into account. A system biology approach should be used in order to systemize all experimental data for rigorous analysis, with the aim of understanding intrinsic pathways of homeostasis, in order to define precise medicine therapy and drug design.

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Gp29 LysA of mycobacteriophage TM4 can hydrolyze peptidoglycan through an N-acetyl-muramoyl-L-alanine amidase activity

Cited by 4 publications

References 48 publications

Investigating Novel Streptomyces Bacteriophage Endolysins as Potential Antimicrobial Agents

Investigating Novel Streptomyces Bacteriophage Endolysins as Potential Antimicrobial Agents

Metagenomic analysis of hot spring soil for mining a novel thermostable enzybiotic

Regulation of Immune Homeostasis via Muramyl Peptides-Low Molecular Weight Bioregulators of Bacterial Origin

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