Bacteriophages Infecting Lactic Acid Bacteria

Murphy, James M.; Mahony, Jennifer; Fitzgerald, Gerald F.; Sinderen, Douwe van

doi:10.1016/b978-0-12-417012-4.00010-7

Cited by 13 publications

(20 citation statements)

References 238 publications

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“…Transmission electron micrographs displayed phage particles with an isometric head (diameter: 55.6 ± 3.2 nm), a neck passage with thin whiskers, a non-contractile flexible tail (length counting in the baseplate: 173.6 ± 5.6 nm; width: 12.0 ± 0.5 nm) and a characteristic double-disc baseplate (length: 12.2 ± 1.0 nm; width: 20.2 ± 1.4 nm) which culminates in short flexible appendages with tiny terminal globular structures. Similar flexible globular structures and capsid-tail dimensions have been observed for other Lactobacillus phages but their exact role remains unknown 3,40 . Interestingly, in some Leuconostoc mesenteroides phages similar baseplate appendages could also adsorb to other parts of the tail in an inconsistent manner 41 .…”

Section: Phage Morphologysupporting

confidence: 54%

“…S3-S5 online). On the whole, such a pattern of genome organisation corresponds to most Lactobacillus phage isolates 3 . Most of the proteins that could be assigned functions to were identified in all three phages.…”

Section: Description Of Phage Modulesmentioning

confidence: 94%

“…Moreover, some benefits of those lactic acid bacteria recognised as probiotics entail health-promoting effects 2 . Lactic acid bacteria are encompassed within the genera of Lactococcus, Pediococcus, Streptococcus, Enterococcus, Oenococcus, Leuconostoc, Lactobacillus, Fructobacillus, Weissella and Carnobacterium [3][4][5] .A member of the genus Lactobacillus, L. plantarum is a versatile lactic acid bacterium of great potential for the food industry. This is due to the increasing popularity of L. plantarum as starter and adjunct culture from dairy to wine fermentations 6,7 .…”

Section: Introductionmentioning

confidence: 99%

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Isolation and characterisation of novel phages infectingLactobacillus plantarumand proposal of a new genus, “Silenusvirus”

Kyrkou

Carstens

Ellegaard-Jensen

et al. 2019

Preprint

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Bacteria of Lactobacillus sp. are very useful to humans. However, the biology and genomic diversity of their (bacterio)phage enemies remains understudied. Knowledge on Lactobacillus phage diversity should broaden to develop efficient phage control strategies. To this end, organic waste samples were screened for phages against two wine-related Lactobacillus plantarum strains. Isolates were shotgun sequenced and compared against the phage database and each other by phylogenetics and comparative genomics. The new isolates had only three distant relatives from the database but displayed a high overall degree of genomic similarity amongst them. The latter allowed for the use of one isolate as a representative to conduct transmission electron microscopy and structural protein sequencing, and to study phage adsorption and growth kinetics. The microscopy and proteomics tests confirmed the observed diversity of the new isolates and supported their classification to the family Siphoviridae and the proposal of the new phage genus "Silenusvirus". Methods Environmental samples, phage assays and bacterial strainsLactobacillus phages were isolated from organic household waste samples. The samples were collected from two different organic waste treatment plants in Denmark (treatment plants A and B). Initially, the samples were split into two subsamples and processed as detailed in Kyrkou et al 12 . The resulting filtrates were screened for phages using the double agar overlay method 16 and a top layer of 0.4% w/v agarose. Specifically, efficiency of plating assays were performed against two indicator strains that had earlier been acquired from private collections, L. plantarum L1 (wine fermentation isolate) and L. plantarum MW-1 (grape isolate). Single plaques were resuspended in 0.7 mL SM buffer and later filtered by 0.45-µm pore size PVDF spin filters (Ciro, Florida, USA). Each purified plaque underwent two further reisolation-filtration cycles to ensure pure stock cultures and was stored at 4°C. For transmission electron microscopy (TEM) and protein sequencing, lysates of 10 10 plaque-forming units (PFUs)/mL) were further purified and concentrated using caesium chloride (CsCl) gradient ultracentrifugation, as described elsewhere 17 . All incubations of phage manipulations were done at 25°C using De Man, Rogosa and Sharpe (MRS) broth and agar media supplemented with 10 mM CaCl 2 (MRSΦ), whereas indicator strains were grown in MRS at 37°C. TEM analysis and structural protein sequencingPhage morphology and structural proteins were determined using the CsCl-purified stocks. Micrographs of phage Silenus were generated as in other studies 12,18 . The mean values and standard deviations of all Silenus virion dimensions were elucidated after inspection of 20-23 phage particles. Structural proteins were sequenced following published protocols 12,18 . Briefly, 100 µL of the CsCl-purified stocks were filtered through an Amicon Ultra filter unit (MWCO 30k Da) and desalted four times. Phage particles (10 µL) were denaturised in 6 M urea, ...

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Section: Phage Morphologysupporting

confidence: 54%

Section: Description Of Phage Modulesmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

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Isolation and characterisation of novel phages infectingLactobacillus plantarumand proposal of a new genus, “Silenusvirus”

Kyrkou

Carstens

Ellegaard-Jensen

et al. 2019

Preprint

View full text Add to dashboard Cite

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“…Bacteriophage (phage) attacks often hamper their activity and by extend cause economic losses because of fermentation failure or poor product quality [8]. Owing to the acute threat they pose to industrially-relevant starters, Lactococcus and Streptococcus phages have been long and extensively studied [9], [10], [11]. On the other hand, the knowledge gaps on the biology of Lactobacillus phages and their genetic heterogeneity have negatively impacted taxonomic efforts [12], [13].…”

Section: Introductionmentioning

confidence: 99%

Expanding the Diversity of Myoviridae Phages Infecting Lactobacillus Plantarum - A Novel Lineage of Lactobacillus Phages Comprising Five New Members

Kyrkou

Carstens

Ellegaard-Jensen

et al. 2019

Preprint

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Lactobacillus plantarum is a bacterium with promising applications to the food industry and agriculture and probiotic properties. So far, bacteriophages of this bacterium have been moderately addressed. We examined the diversity of five new L. plantarum phages via whole genome shotgun sequencing and in silico protein predictions. Moreover, we looked into their phylogeny and their potential genomic similarities to other complete phage genome records through extensive nucleotide and protein comparisons. These analyses revealed a high degree of similarity among the five phages, which extended to the vast majority of predicted virion-associated proteins. Based on these, we selected one of the phages as a representative and performed transmission electron microscopy and structural protein sequencing tests. Overall, the results suggested that the five phages belong to the family Myoviridae, they have a long genome of 137.973-141.344 bp, a G/C content of 36,3-36,6% that is quite distinct from their host’s, and, surprisingly, seven to 15 tRNAs. Only an average 41/174 of their predicted genes were assigned a function. The comparative analyses unraveled considerable genetic diversity for the five L. plantarum phages of this study. Hence, the new genus “Semelevirus” was proposed, which comprises exclusively the five phages. This novel lineage of Lactobacillus phages provides further insight into the genetic heterogeneity of phages infecting Lactobacillus sp.. The five new Lactobacillus phages have a potential value for the development of more robust starters through, for example, the selection of mutants insensitive to phage infections. The five phages could also form part of phage cocktails, which producers would apply in different stages of L. plantarum fermentations in order to create a range of organoleptic outputs.

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“…Bacteriophage (phage) attacks often hamper their activity and by extend cause economic losses because of fermentation failure or poor product quality [7]. Owing to the acute threat they pose to industrially-relevant starters, Lactococcus and Streptococcus phages have been long and extensively studied [8], [9], [10]. On the other hand, the knowledge gaps on the biology of Lactobacillus phages and their genetic heterogeneity have negatively impacted taxonomic efforts [11], [12].…”

Section: Introductionmentioning

confidence: 99%

Expanding the Diversity of Myoviridae Phages Infecting Lactobacillus Plantarum - A Novel Lineage of Lactobacillus Phages Comprising Five New Members

Kyrkou¹,

Carstens²,

Ellegaard-Jensen³

et al. 2018

Preprint

View full text Add to dashboard Cite

Lactobacillus plantarum is a bacterium with promising applications to the food industry and agriculture and probiotic properties. So far, bacteriophages of this bacterium have been moderately addressed. We examined the morphology and diversity of five new L. plantarum phages via transmission electron microscopy, whole genome shotgun sequencing and in silico protein predictions. Moreover, we looked into their phylogeny and their potential genomic similarities to other complete phage genome records through extensive nucleotide and protein comparisons. The five phages share a high degree of similarity among them and belong to the family Myoviridae. They have a long genome of 137.973-141.344 bp, a G/C content of 36,3-36,6% that is quite distinct from their host’s, and, surprisingly, seven to 15 tRNAs. Only an average 35/174 of their predicted genes were assigned a function. Overall, the comparative analyses unraveled considerable genetic diversity for our five L. plantarum phages. Hence, the new genus “Semelevirus” was proposed, which comprises exclusively the five phages. This novel lineage of Lactobacillus phages provides further insight into the genetic heterogeneity of phages infecting Lactobacillus sp. Our results on Lactobacillus phages have a potential value for the development of anti-phage strategies as well as for the manipulation of L. plantarum fermentations.

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Bacteriophages Infecting Lactic Acid Bacteria

Cited by 13 publications

References 238 publications

Isolation and characterisation of novel phages infectingLactobacillus plantarumand proposal of a new genus, “Silenusvirus”

Isolation and characterisation of novel phages infectingLactobacillus plantarumand proposal of a new genus, “Silenusvirus”

Expanding the Diversity of Myoviridae Phages Infecting Lactobacillus Plantarum - A Novel Lineage of Lactobacillus Phages Comprising Five New Members

Expanding the Diversity of Myoviridae Phages Infecting Lactobacillus Plantarum - A Novel Lineage of Lactobacillus Phages Comprising Five New Members

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