Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity
Staphylococcus aureus causes various infections in humans and animals, the skin being the principal reservoir of this pathogen. The widespread occurrence of methicillin-resistant S. aureus (MRSA) limits the elimination and treatment of this pathogen. Phage lytic proteins have been proven as efficien...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.723834/full |
| _version_ | 1818671276817907712 |
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| author | Diana Gutiérrez Diana Gutiérrez Diana Gutiérrez Lorena Rodríguez-Rubio Patricia Ruas-Madiedo Patricia Ruas-Madiedo Lucía Fernández Lucía Fernández Ana Belén Campelo Yves Briers Martin Weiss Nielsen Karl Pedersen Rob Lavigne Pilar García Pilar García Ana Rodríguez Ana Rodríguez |
| author_facet | Diana Gutiérrez Diana Gutiérrez Diana Gutiérrez Lorena Rodríguez-Rubio Patricia Ruas-Madiedo Patricia Ruas-Madiedo Lucía Fernández Lucía Fernández Ana Belén Campelo Yves Briers Martin Weiss Nielsen Karl Pedersen Rob Lavigne Pilar García Pilar García Ana Rodríguez Ana Rodríguez |
| author_sort | Diana Gutiérrez |
| collection | DOAJ |
| description | Staphylococcus aureus causes various infections in humans and animals, the skin being the principal reservoir of this pathogen. The widespread occurrence of methicillin-resistant S. aureus (MRSA) limits the elimination and treatment of this pathogen. Phage lytic proteins have been proven as efficient antimicrobials against S. aureus. Here, a set of 12 engineered proteins based on endolysins were conceptualized to select the most optimal following a stepwise funnel approach assessing parameters including turbidity reduction, minimum inhibitory concentration (MIC), time-kill curves, and antibiofilm assays, as well as testing their stability in a broad range of storage conditions (pH, temperature, and ionic strength). The engineered phage lysins LysRODIΔAmi and ClyRODI-H5 showed the highest specific lytic activity (5 to 50 times higher than the rest), exhibited a shelf-life up to 6 months and remained stable at temperatures up to 50°C and in a pH range from 3 to 9. LysRODIΔAmi showed the lower MIC values against all staphylococcal strains tested. Both proteins were able to kill 6 log units of the strain S. aureus Sa9 within 5 min and could remove preformed biofilms (76 and 65%, respectively). Moreover, LysRODIΔAmi could prevent biofilm formation at low protein concentrations (0.15–0.6 μM). Due to its enhanced antibiofilm properties, LysRODIΔAmi was selected to effectively remove S. aureus contamination in both intact and disrupted keratinocyte monolayers. Notably, this protein did not demonstrate any toxicity toward human keratinocytes, even at high concentrations (22.1 μM). Finally, a pig skin ex vivo model was used to evaluate treatment of artificially contaminated pig skin using LysRODIΔAmi (16.5 μg/cm2). Following an early reduction of S. aureus, a second dose of protein completely eradicated S. aureus. Overall, our results suggest that LysRODIΔAmi is a suitable candidate as antimicrobial agent to prevent and treat staphylococcal skin infections. |
| first_indexed | 2024-12-17T07:21:26Z |
| format | Article |
| id | doaj.art-8d280d8d61ed4637a780c397773a7465 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-17T07:21:26Z |
| publishDate | 2021-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-8d280d8d61ed4637a780c397773a74652022-12-21T21:58:45ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-09-011210.3389/fmicb.2021.723834723834Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing ActivityDiana Gutiérrez0Diana Gutiérrez1Diana Gutiérrez2Lorena Rodríguez-Rubio3Patricia Ruas-Madiedo4Patricia Ruas-Madiedo5Lucía Fernández6Lucía Fernández7Ana Belén Campelo8Yves Briers9Martin Weiss Nielsen10Karl Pedersen11Rob Lavigne12Pilar García13Pilar García14Ana Rodríguez15Ana Rodríguez16Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, SpainInstituto de Investigación Sanitaria del Principado de Asturias, Oviedo, SpainLaboratory of Applied Biotechnology, Department of Biotechnology, Ghent University, Ghent, BelgiumLaboratory of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Leuven, BelgiumInstituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, SpainInstituto de Investigación Sanitaria del Principado de Asturias, Oviedo, SpainInstituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, SpainInstituto de Investigación Sanitaria del Principado de Asturias, Oviedo, SpainInstituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, SpainLaboratory of Applied Biotechnology, Department of Biotechnology, Ghent University, Ghent, BelgiumDepartment of Microbiology and Production, National Food Institute, Technical University of Denmark, Lyngby, DenmarkDepartment of Microbiology and Production, National Food Institute, Technical University of Denmark, Lyngby, DenmarkLaboratory of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Leuven, BelgiumInstituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, SpainInstituto de Investigación Sanitaria del Principado de Asturias, Oviedo, SpainInstituto de Productos Lácteos de Asturias (IPLA-CSIC), Asturias, SpainInstituto de Investigación Sanitaria del Principado de Asturias, Oviedo, SpainStaphylococcus aureus causes various infections in humans and animals, the skin being the principal reservoir of this pathogen. The widespread occurrence of methicillin-resistant S. aureus (MRSA) limits the elimination and treatment of this pathogen. Phage lytic proteins have been proven as efficient antimicrobials against S. aureus. Here, a set of 12 engineered proteins based on endolysins were conceptualized to select the most optimal following a stepwise funnel approach assessing parameters including turbidity reduction, minimum inhibitory concentration (MIC), time-kill curves, and antibiofilm assays, as well as testing their stability in a broad range of storage conditions (pH, temperature, and ionic strength). The engineered phage lysins LysRODIΔAmi and ClyRODI-H5 showed the highest specific lytic activity (5 to 50 times higher than the rest), exhibited a shelf-life up to 6 months and remained stable at temperatures up to 50°C and in a pH range from 3 to 9. LysRODIΔAmi showed the lower MIC values against all staphylococcal strains tested. Both proteins were able to kill 6 log units of the strain S. aureus Sa9 within 5 min and could remove preformed biofilms (76 and 65%, respectively). Moreover, LysRODIΔAmi could prevent biofilm formation at low protein concentrations (0.15–0.6 μM). Due to its enhanced antibiofilm properties, LysRODIΔAmi was selected to effectively remove S. aureus contamination in both intact and disrupted keratinocyte monolayers. Notably, this protein did not demonstrate any toxicity toward human keratinocytes, even at high concentrations (22.1 μM). Finally, a pig skin ex vivo model was used to evaluate treatment of artificially contaminated pig skin using LysRODIΔAmi (16.5 μg/cm2). Following an early reduction of S. aureus, a second dose of protein completely eradicated S. aureus. Overall, our results suggest that LysRODIΔAmi is a suitable candidate as antimicrobial agent to prevent and treat staphylococcal skin infections.https://www.frontiersin.org/articles/10.3389/fmicb.2021.723834/fullendolysinprotein engineeringantimicrobialStaphylococcus aureusskin decontamination |
| spellingShingle | Diana Gutiérrez Diana Gutiérrez Diana Gutiérrez Lorena Rodríguez-Rubio Patricia Ruas-Madiedo Patricia Ruas-Madiedo Lucía Fernández Lucía Fernández Ana Belén Campelo Yves Briers Martin Weiss Nielsen Karl Pedersen Rob Lavigne Pilar García Pilar García Ana Rodríguez Ana Rodríguez Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity Frontiers in Microbiology endolysin protein engineering antimicrobial Staphylococcus aureus skin decontamination |
| title | Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity |
| title_full | Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity |
| title_fullStr | Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity |
| title_full_unstemmed | Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity |
| title_short | Design and Selection of Engineered Lytic Proteins With Staphylococcus aureus Decolonizing Activity |
| title_sort | design and selection of engineered lytic proteins with staphylococcus aureus decolonizing activity |
| topic | endolysin protein engineering antimicrobial Staphylococcus aureus skin decontamination |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.723834/full |
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