Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5
<i>Eggerthella lenta</i> is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having <i>E. lenta</i> as host. The double-layer agar method for isolating phages was adapted to anaerobic conditi...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-07-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/14/8/1598 |
| _version_ | 1797407569238556672 |
|---|---|
| author | 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 |
| author_facet | 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 |
| author_sort | Sabrina Sprotte |
| collection | DOAJ |
| description | <i>Eggerthella lenta</i> is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having <i>E. lenta</i> 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 <i>E. lenta</i> strain of intestinal origin. For this, anaerobically grown <i>E. lenta</i> 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 <i>Siphoviridae</i> 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. |
| first_indexed | 2024-03-09T03:43:27Z |
| format | Article |
| id | doaj.art-69cf68934dda47eba1999e5c366a5227 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T03:43:27Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-69cf68934dda47eba1999e5c366a52272023-12-03T14:37:13ZengMDPI AGViruses1999-49152022-07-01148159810.3390/v14081598Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5Sabrina Sprotte0Torben S. Rasmussen1Gyu-Sung Cho2Erik Brinks3René Lametsch4Horst Neve5Finn K. Vogensen6Dennis S. Nielsen7Charles M. A. P. Franz8Department of Microbiology and Biotechnology, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 24103 Kiel, GermanyDepartment of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, DenmarkDepartment of Microbiology and Biotechnology, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 24103 Kiel, GermanyDepartment of Microbiology and Biotechnology, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 24103 Kiel, GermanyDepartment of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, DenmarkDepartment of Microbiology and Biotechnology, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 24103 Kiel, GermanyDepartment of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, DenmarkDepartment of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, DenmarkDepartment of Microbiology and Biotechnology, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 24103 Kiel, Germany<i>Eggerthella lenta</i> is a common member of the human gut microbiome. We here describe the isolation and characterization of a putative virulent bacteriophage having <i>E. lenta</i> 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 <i>E. lenta</i> strain of intestinal origin. For this, anaerobically grown <i>E. lenta</i> 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 <i>Siphoviridae</i> 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.https://www.mdpi.com/1999-4915/14/8/1598anaerobe<i>Eggerthella lenta</i>virulent phage<i>Siphoviridae</i>genome sequence |
| spellingShingle | 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 Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5 Viruses anaerobe <i>Eggerthella lenta</i> virulent phage <i>Siphoviridae</i> genome sequence |
| title | Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5 |
| title_full | Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5 |
| title_fullStr | Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5 |
| title_full_unstemmed | Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5 |
| title_short | Morphological and Genetic Characterization of <i>Eggerthella lenta</i> Bacteriophage PMBT5 |
| title_sort | morphological and genetic characterization of i eggerthella lenta i bacteriophage pmbt5 |
| topic | anaerobe <i>Eggerthella lenta</i> virulent phage <i>Siphoviridae</i> genome sequence |
| url | https://www.mdpi.com/1999-4915/14/8/1598 |
| work_keys_str_mv | AT sabrinasprotte morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT torbensrasmussen morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT gyusungcho morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT erikbrinks morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT renelametsch morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT horstneve morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT finnkvogensen morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT dennissnielsen morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 AT charlesmapfranz morphologicalandgeneticcharacterizationofieggerthellalentaibacteriophagepmbt5 |