The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing
The rising antimicrobial resistance is particularly alarming for <i>Acinetobacter baumannii</i>, calling for the discovery and evaluation of alternatives to treat <i>A. baumannii</i> infections. Some bacteriophages produce a structural protein that depolymerizes capsular exop...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-05-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/11/5/677 |
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| author | Karim Abdelkader Diana Gutiérrez Agnieszka Latka Dimitri Boeckaerts Zuzanna Drulis-Kawa Bjorn Criel Hans Gerstmans Amal Safaan Ahmed S. Khairalla Yasser Gaber Tarek Dishisha Yves Briers |
| author_facet | Karim Abdelkader Diana Gutiérrez Agnieszka Latka Dimitri Boeckaerts Zuzanna Drulis-Kawa Bjorn Criel Hans Gerstmans Amal Safaan Ahmed S. Khairalla Yasser Gaber Tarek Dishisha Yves Briers |
| author_sort | Karim Abdelkader |
| collection | DOAJ |
| description | The rising antimicrobial resistance is particularly alarming for <i>Acinetobacter baumannii</i>, calling for the discovery and evaluation of alternatives to treat <i>A. baumannii</i> infections. Some bacteriophages produce a structural protein that depolymerizes capsular exopolysaccharide. Such purified depolymerases are considered as novel antivirulence compounds. We identified and characterized a depolymerase (DpoMK34) from Acinetobacter phage vB_AbaP_PMK34 active against the clinical isolate <i>A. baumannii</i> MK34. In silico analysis reveals a modular protein displaying a conserved N-terminal domain for anchoring to the phage tail, and variable central and C-terminal domains for enzymatic activity and specificity. AlphaFold-Multimer predicts a trimeric protein adopting an elongated structure due to a long α-helix, an enzymatic β-helix domain and a hypervariable 4 amino acid hotspot in the most ultimate loop of the C-terminal domain. In contrast to the tail fiber of phage T3, this hypervariable hotspot appears unrelated with the primary receptor. The functional characterization of DpoMK34 revealed a mesophilic enzyme active up to 50 °C across a wide pH range (4 to 11) and specific for the capsule of <i>A. baumannii</i> MK34. Enzymatic degradation of the <i>A. baumannii</i> MK34 capsule causes a significant drop in phage adsorption from 95% to 9% after 5 min. Although lacking intrinsic antibacterial activity, DpoMK34 renders <i>A. baumannii</i> MK34 fully susceptible to serum killing in a serum concentration dependent manner. Unlike phage PMK34, DpoMK34 does not easily select for resistant mutants either against PMK34 or itself. In sum, DpoMK34 is a potential antivirulence compound that can be included in a depolymerase cocktail to control difficult to treat <i>A. baumannii</i> infections. |
| first_indexed | 2024-03-10T03:27:14Z |
| format | Article |
| id | doaj.art-5475799332b641e4b5f97f29d50649a1 |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-10T03:27:14Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-5475799332b641e4b5f97f29d50649a12023-11-23T09:49:33ZengMDPI AGAntibiotics2079-63822022-05-0111567710.3390/antibiotics11050677The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum KillingKarim Abdelkader0Diana Gutiérrez1Agnieszka Latka2Dimitri Boeckaerts3Zuzanna Drulis-Kawa4Bjorn Criel5Hans Gerstmans6Amal Safaan7Ahmed S. Khairalla8Yasser Gaber9Tarek Dishisha10Yves Briers11Department of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumDepartment of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumDepartment of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumDepartment of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumDepartment of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wroclaw, Przybyszewskiego 63, 51-148 Wrocław, PolandDepartment of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumDepartment of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumDepartment of Microbiology and Immunology, Faculty of Pharmacy, Menoufia University, Shebin El-Koum 51132, EgyptDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62511, EgyptDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62511, EgyptDepartment of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62511, EgyptDepartment of Biotechnology, Ghent University, Valentin Vaerwyckweg 1, 9000 Gent, BelgiumThe rising antimicrobial resistance is particularly alarming for <i>Acinetobacter baumannii</i>, calling for the discovery and evaluation of alternatives to treat <i>A. baumannii</i> infections. Some bacteriophages produce a structural protein that depolymerizes capsular exopolysaccharide. Such purified depolymerases are considered as novel antivirulence compounds. We identified and characterized a depolymerase (DpoMK34) from Acinetobacter phage vB_AbaP_PMK34 active against the clinical isolate <i>A. baumannii</i> MK34. In silico analysis reveals a modular protein displaying a conserved N-terminal domain for anchoring to the phage tail, and variable central and C-terminal domains for enzymatic activity and specificity. AlphaFold-Multimer predicts a trimeric protein adopting an elongated structure due to a long α-helix, an enzymatic β-helix domain and a hypervariable 4 amino acid hotspot in the most ultimate loop of the C-terminal domain. In contrast to the tail fiber of phage T3, this hypervariable hotspot appears unrelated with the primary receptor. The functional characterization of DpoMK34 revealed a mesophilic enzyme active up to 50 °C across a wide pH range (4 to 11) and specific for the capsule of <i>A. baumannii</i> MK34. Enzymatic degradation of the <i>A. baumannii</i> MK34 capsule causes a significant drop in phage adsorption from 95% to 9% after 5 min. Although lacking intrinsic antibacterial activity, DpoMK34 renders <i>A. baumannii</i> MK34 fully susceptible to serum killing in a serum concentration dependent manner. Unlike phage PMK34, DpoMK34 does not easily select for resistant mutants either against PMK34 or itself. In sum, DpoMK34 is a potential antivirulence compound that can be included in a depolymerase cocktail to control difficult to treat <i>A. baumannii</i> infections.https://www.mdpi.com/2079-6382/11/5/677capsuledepolymerasephage<i>Acinetobacter baumannii</i>tailspikeantivirulence |
| spellingShingle | Karim Abdelkader Diana Gutiérrez Agnieszka Latka Dimitri Boeckaerts Zuzanna Drulis-Kawa Bjorn Criel Hans Gerstmans Amal Safaan Ahmed S. Khairalla Yasser Gaber Tarek Dishisha Yves Briers The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing Antibiotics capsule depolymerase phage <i>Acinetobacter baumannii</i> tailspike antivirulence |
| title | The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing |
| title_full | The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing |
| title_fullStr | The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing |
| title_full_unstemmed | The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing |
| title_short | The Specific Capsule Depolymerase of Phage PMK34 Sensitizes <i>Acinetobacter baumannii</i> to Serum Killing |
| title_sort | specific capsule depolymerase of phage pmk34 sensitizes i acinetobacter baumannii i to serum killing |
| topic | capsule depolymerase phage <i>Acinetobacter baumannii</i> tailspike antivirulence |
| url | https://www.mdpi.com/2079-6382/11/5/677 |
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