High cell-density fermentation, expression and purification of bacteriophage lysin TSPphg, a thermostable antimicrobial protein from extremophilic Thermus bacteriophage TSP4

Wu, Feng; Zhang, Guanling; Peng, Jiani; Ji, Xinyu; Hai, Jun; Deng, Xian-Yu; Lü, Lin

doi:10.1016/j.pep.2020.105676

Cited by 5 publications

(3 citation statements)

References 43 publications

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“…In addition, we frequently observed that the bactericidal effect of the lytic enzyme slowly decreased with increasing storage duration in the freezer. Indeed, low stability and an inactivation tendency are major shortcomings of protein antibacterial agents ( Wang et al, 2020b ). These shortcomings should be the focus of future research, and efforts should also be made to improve the stability and storage life of these agents.…”

Section: Discussionmentioning

confidence: 99%

Bacteriophage Lytic Enzyme P9ly as an Alternative Antibacterial Agent Against Antibiotic-Resistant Shigella dysenteriae and Staphylococcus aureus

Feng

Xiao

et al. 2022

Front. Microbiol.

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Developing new strategies to replace or supplement antibiotics to combat bacterial infection is a pressing task in the field of microbiological research. In this study, we report a lytic enzyme named P9ly deriving from the bacteriophage PSD9 that could infect multidrug-resistant Shigella. This enzyme was identified through whole-genome sequencing of PSD9. The results show that P9ly contains a conserved T4-like_lys domain and belongs to the phage lysozyme family. Recombinant P9ly obtained from protein purification presented biological activity and could digest bacterial cell walls (CW), resulting in the destruction of cell structure and leakage of intracellular components. Furthermore, P9ly exhibited bacteriolytic and bactericidal activity on different strains, especially multidrug-resistant Gram-negative Shigella dysenteriae and Gram-positive Staphylococcus aureus. Additionally, combined use of P9ly with ceftriaxone sodium (CRO) could decrease necessary dose of the antibiotic used and improve the antibacterial effect. In summary, under the current backdrop of extensive antibiotic usage and the continuous emergence of bacterial resistance, this study provides an insight into developing bacteriophage-based antibacterial agents against both Gram-negative and Gram-positive pathogens.

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Section: Discussionmentioning

confidence: 99%

Bacteriophage Lytic Enzyme P9ly as an Alternative Antibacterial Agent Against Antibiotic-Resistant Shigella dysenteriae and Staphylococcus aureus

Feng

Xiao

et al. 2022

Front. Microbiol.

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“…The clpB gene was expressed in E. coli BL21 (DE3), and the ClpB protein was purified as a hexamer as previously reported ( Lupoli et al, 2016 ; Rizo et al, 2019 ). In brief, E. coli BL21 (DE3)- clpB was incubated in LB medium for heterologous gene expression at 37°C, supplemented with 0.5 mM isopropyl-β-D-thiogalactoside, and cultivated for 12 h. The proteins were harvested and analyzed by SDS-PAGE ( Wang et al, 2020 ). Whole cells were disrupted using sonication ( Wang et al, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

“…In brief, E. coli BL21 (DE3)-clpB was incubated in LB medium for heterologous gene expression at 37 • C, supplemented with 0.5 mM isopropyl-β-D-thiogalactoside, and cultivated for 12 h. The proteins were harvested and analyzed by SDS-PAGE (Wang et al, 2020). Whole cells were disrupted using sonication (Wang et al, 2020). Subsequently, the recombinant protein was purified using Ni-NTA affinity chromatography (Sangon Biotech, China).…”

Section: Construction Of Plasmids and Strainsmentioning

confidence: 99%

Role of ClpB From Corynebacterium crenatum in Thermal Stress and Arginine Fermentation

et al. 2020

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ClpB, an ATP-dependent molecular chaperone, is involved in metabolic pathways and plays important roles in microorganisms under stress conditions. Metabolic pathways and stress resistance are important characteristics of industrially-relevant bacteria during fermentation. Nevertheless, ClpB-related observations have been rarely reported in industrially-relevant microorganisms. Herein, we found a homolog of ClpB from Corynebacterium crenatum. The amino acid sequence of ClpB was analyzed, and the recombinant ClpB protein was purified and characterized. The full function of ClpB requires DnaK as chaperone protein. For this reason, dnaK/clpB deletion mutants and the complemented strains were constructed to investigate the role of ClpB. The results showed that DnaK/ClpB is not essential for the survival of C. crenatum MT under pH and alcohol stresses. The ClpB-deficient or DnaK-deficient C. crenatum mutants showed weakened growth during thermal stress. In addition, the results demonstrated that deletion of the clpB gene affected glucose consumption and L-arginine, L-glutamate, and lactate production during fermentation.

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A novel bacteriocin against multiple foodborne pathogens from Lacticaseibacillus rhamnosus isolated from juice ferments: ATF perfusion-based preparation of viable cells, characterization, antibacterial and antibiofilm activity

Chen¹,

Yang²,

Ci³

et al. 2023

Current Research in Food Science

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High cell-density fermentation, expression and purification of bacteriophage lysin TSPphg, a thermostable antimicrobial protein from extremophilic Thermus bacteriophage TSP4

Cited by 5 publications

References 43 publications

Bacteriophage Lytic Enzyme P9ly as an Alternative Antibacterial Agent Against Antibiotic-Resistant Shigella dysenteriae and Staphylococcus aureus

Bacteriophage Lytic Enzyme P9ly as an Alternative Antibacterial Agent Against Antibiotic-Resistant Shigella dysenteriae and Staphylococcus aureus

Role of ClpB From Corynebacterium crenatum in Thermal Stress and Arginine Fermentation

A novel bacteriocin against multiple foodborne pathogens from Lacticaseibacillus rhamnosus isolated from juice ferments: ATF perfusion-based preparation of viable cells, characterization, antibacterial and antibiofilm activity

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