Morphological and Genetic Characterization of Eggerthella lenta Bacteriophage PMBT5

Sabrina Sprotte*, Torben S. Rasmussen, Gyu-Sung Cho, Erik Brinks, René Lametsch, Horst Neve, Finn K. Vogensen, Dennis S. Nielsen, Charles M.A.P. Franz

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Eggerthella lenta is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having E. lenta as host. The double-layer agar method for isolating phages was adapted to anaerobic conditions for isolating bacteriophage PMBT5 from sewage on a strictly anaerobic E. lenta strain of intestinal origin. For this, anaerobically grown E. lenta cells were concentrated by centrifugation and used for a 24 h phage enrichment step. Subsequently, this suspension was added to anaerobically prepared top (soft) agar in Hungate tubes and further used in the double-layer agar method. Based on morphological characteristics observed by transmission electron microscopy, phage PMBT5 could be assigned to the Siphoviridae phage family. It showed an isometric head with a flexible, noncontractile tail and a distinct single 45 nm tail fiber under the baseplate. Genome sequencing and assembly resulted in one contig of 30,930 bp and a mol% GC content of 51.3, consisting of 44 predicted protein-encoding genes. Phage-related proteins could be largely identified based on their amino acid sequence, and a comparison with metagenomes in the human virome database showed that the phage genome exhibits similarity to two distantly related phages.

Original languageEnglish
Article number1598
JournalViruses
Volume14
Issue number8
ISSN1999-4915
DOIs
Publication statusPublished - 2022

Bibliographical note

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© 2022 by the authors.

Keywords

  • anaerobe
  • Eggerthella lenta
  • genome sequence
  • Siphoviridae
  • virulent phage

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