Characterization and adsorption of Lactobacillus virulent phage P1

Chen, X.; Yang, Xi; Zhang, H.; Wang, Z.; Fan, Mingtao; Liu, Y.; Wu, W.

doi:10.3168/jds.2016-11332

Cited by 21 publications

(21 citation statements)

References 16 publications

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“…A major difference between phages P1 and P2 was size. Phage P1 was larger, with an isometric capsid of 71.7 ± 3.0 nm and long noncontractile tails (approximately 272 ± 3.0 nm long and 11.3 ± 1.5 nm wide; Chen et al, 2016). Ackermann (2007) also reported that for various Lactobacillus phages, almost 60% belonged to the Siphoviridae family.…”

Section: Electron Microscopymentioning

confidence: 97%

“…Nonspecific adsorption between a phage and the bacterial surface is reversible, and external conditions such as temperature and pH are influential. Studies concerning the physiological and environmental factors on phage propagation and adsorption have been previously reported (Capra et al, 2006;Briggiler Marcó et al, 2010;Cvirkaitė-Krupovič et al, 2010;Mahony et al, 2015;Chen et al, 2016). In the event that the bacteriophage tail and bacterial surface receptor sites come together due to specific covalent binding, an irreversible adsorption process ensues.…”

Section: Introductionmentioning

confidence: 99%

“…Lactobacillus plantarum IMAU 10120 is a starter culture that exhibits several desirable properties, such as high acid and bile tolerance, good aggregation, antibacterial activities, high soy milk fermentation efficiency, and strong stability upon storage. Chen et al (2016) isolated a Lactobacillus virulent phage (P1) from its abnormal fermented liquid and researched its biological properties and the influence of physicochemical parameters on its adsorption. Unfortunately, the abnormal fermentation occurred again, and a new bacteriophage (P2) was isolated.…”

Section: Introductionmentioning

confidence: 99%

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Characterization and adsorption of a Lactobacillus plantarum virulent phage

Chen

Guo

Liu

et al. 2019

Journal of Dairy Science

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Bacteriophage infection of lactic acid bacteria is considered one of the biggest worldwide problems in the food industry. Bacteriophages may cause negative effects on the fermentation of various dairy-based products. A virulent bacteriophage was isolated from an abnormal fermentation liquid of Lactobacillus plantarum IMAU10120. The characterization and influence of temperature, pH, divalent cations, and chloramphenicol on the adsorption ability of this phage were evaluated. The results showed that this phage belonged to the Siphoviridae family. It exhibited a burst time of 135 min and a burst size of approximately 215 counts expressed per milliliter per infective center. No significant effect was shown to influence its viability and adsorption at 10 to 37°C. More than 90% of phages exhibited infectivity from pH 5 to 9. Divalent ions and chloramphenicol did not have a significant influence on the adsorption of this phage. The information obtained in this study will enrich the database of lactobacilli virulent phages and provide a basis of information for the control of phages in the food fermentation industry.

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Section: Electron Microscopymentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization and adsorption of a Lactobacillus plantarum virulent phage

Chen

Guo

Liu

et al. 2019

Journal of Dairy Science

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“…Since TP-84 dissociates in the presence of EDTA [ 15 ], one could assume that divalent ions, particularly Ca 2+ , are involved in the stabilization of the virions. Furthermore, many bacteriophages require calcium ions for infectivity [ 25 , 26 ]. Since many calcium-binding proteins share an EF-hand motif [ 27 ], we screened the TP-84 putative structural proteins in Prosite [ 28 ] for the PDOC00018 signature sequence.…”

Section: Resultsmentioning

confidence: 99%

Sequence, genome organization, annotation and proteomics of the thermophilic, 47.7-kb Geobacillus stearothermophilus bacteriophage TP-84 and its classification in the new Tp84virus genus

et al. 2018

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Bacteriophage TP-84 is a well-characterized bacteriophage of historical interest. It is a member of the Siphoviridae, and infects a number of thermophilic Geobacillus (Bacillus) stearothermophilus strains. Its’ 47.7-kbp double-stranded DNA genome revealed the presence of 81 coding sequences (CDSs) coding for polypeptides of 4 kDa or larger. Interestingly, all CDSs are oriented in the same direction, pointing to a dominant transcription direction of one DNA strand. Based on a homology search, a hypothetical function could be assigned to 31 CDSs. No RNA or DNA polymerase-coding genes were found on the bacteriophage genome indicating that TP-84 relies on the host’s transcriptional and replication enzymes. The TP84 genome is tightly packed with CDSs, typically spaced by several-to-tens of bp or often overlapping. The genome contains five putative promoter-like sequences showing similarity to the host promoter consensus sequence and allowing for a 2-bp mismatch. In addition, ten putative rho-independent terminators were detected. Because the genome sequence shows essentially no similarity to any previously characterised bacteriophage, TP-84 should be considered a new species in an undefined genus within the Siphoviridae family. Thus a taxonomic proposal of a new Tp84virus genus has been accepted by the International Committee on Taxonomy of Viruses. The bioinformatics genome analysis was verified by confirmation of 33 TP-84 proteins, which included: a) cloning of a selected CDS in Escherichia coli, coding for a DNA single-stranded binding protein (SSB; gene TP84_63), b) purification and functional assays of the recombinant TP-84 SSB, which has been shown to improve PCR reactions, c) mass spectrometric (MS) analysis of TP-84 bacteriophage capsid proteins, d) purification of TP-84 endolysin activity, e) MS analysis of the host cells from infection time course.

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“…Since little is known about the pH stability of E. faecalis phages, we compared the pH stability of phage HEf13 with those of other Gram-positive bacteria phages. Phage HEf13 titers were not affected by incubation at pH 11 and remained at around 30% at pH 12 ( Figure 4B), whereas reported titers of other phages were almost completely abolished at pH 11 (Li and Zhang, 2014;Chen et al, 2016;Phothichaisri et al, 2018). These comparisons demonstrated that phage HEf13 is relatively more stable to high temperatures and strongly basic conditions than other phages.…”

Section: Discussionmentioning

confidence: 83%

The Novel Enterococcus Phage vB_EfaS_HEf13 Has Broad Lytic Activity Against Clinical Isolates of Enterococcus faecalis

Lee

et al. 2019

Front. Microbiol.

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Enterococcus faecalis is a Gram-positive, facultative anaerobic bacterium frequently found in the gastrointestinal tract, oral cavity, and periodontal tissue. Although it is considered a commensal, it can cause bacteremia, endocarditis, endodontic infections, and urinary tract infections. Because antibiotics are cytotoxic not only to pathogens, but also to health-beneficial commensals, phage therapy has emerged as an alternative strategy to specifically control pathogenic bacteria with minimal damage to the normal flora. In this study, we isolated a novel phage, Enterococcus phage vB_EfaS_HEf13 (phage HEf13), with broad lytic activity against 12 strains of E. faecalis among the three laboratory strains and 14 clinical isolates of E. faecalis evaluated. Transmission electron microscopy showed that phage HEf13 has morphological characteristics of the family Siphoviridae. Phage HEf13 was stable at a wide range of temperature (4-60 • C) and showed tolerance to acid or alkaline (pH 3-12) growth conditions. Phage HEf13 had a short latent period (25 min) with a large burst size (approximately 352 virions per infected cell). The lytic activity of phage HEf13 at various multiplicities of infection consistently inhibited the growth of diverse clinical isolates of E. faecalis without any lysogenic process. Moreover, phage HEf13 showed an effective lytic activity against E. faecalis on human dentin ex vivo infection model. Whole genome analysis demonstrated that the phage HEf13 genome contains 57,811 bp of double-stranded DNA with a GC content of 40.1% and 95 predicted open reading frames (ORFs). Annotated functional ORFs were mainly classified into four groups: DNA replication/packaging/regulation, phage structure, host cell lysis, and additional functions such as RNA transcription. Comparative genomic analysis demonstrated that phage HEf13 is a novel phage that belongs to the Sap6virus lineage. Furthermore, the results of multiple sequence alignment showed that polymorphism of phage infection protein of E. faecalis (PIP EF) contributes to determine the host specificity of phage HEf13 against various E. faecalis strains. Collectively, these results suggest that phage HEf13 has characteristics of a lytic phage, and is a potential therapeutic agent for treatment or prevention of E. faecalis-associated infectious diseases.

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Characterization and adsorption of Lactobacillus virulent phage P1

Cited by 21 publications

References 16 publications

Characterization and adsorption of a Lactobacillus plantarum virulent phage

Characterization and adsorption of a Lactobacillus plantarum virulent phage

Sequence, genome organization, annotation and proteomics of the thermophilic, 47.7-kb Geobacillus stearothermophilus bacteriophage TP-84 and its classification in the new Tp84virus genus

The Novel Enterococcus Phage vB_EfaS_HEf13 Has Broad Lytic Activity Against Clinical Isolates of Enterococcus faecalis

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