Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing
In view of the current threat of antibiotic resistance, new antimicrobials with low risk of resistance development are demanded. Lcn972 is a lactococcal bacteriocin that inhibits septum formation by binding to the cell wall precursor lipid II in <i>Lactococcus</i>. It has a species-speci...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-02-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/11/2/501 |
| _version_ | 1797619176881258496 |
|---|---|
| author | Susana Escobedo Ana B. Campelo Özgün C. O. Umu María Jesús López-González Ana Rodríguez Dzung B. Diep Beatriz Martínez |
| author_facet | Susana Escobedo Ana B. Campelo Özgün C. O. Umu María Jesús López-González Ana Rodríguez Dzung B. Diep Beatriz Martínez |
| author_sort | Susana Escobedo |
| collection | DOAJ |
| description | In view of the current threat of antibiotic resistance, new antimicrobials with low risk of resistance development are demanded. Lcn972 is a lactococcal bacteriocin that inhibits septum formation by binding to the cell wall precursor lipid II in <i>Lactococcus</i>. It has a species-specific spectrum of activity, making Lcn972 an attractive template to develop or improve existing antibiotics. The aim of this work was to identify mutations present in the Lcn972-resistant clone <i>Lactococcus cremoris</i> D1-20, previously evolved from the sensitive strain <i>L. cremoris</i> MG1614. Whole-genome sequencing and comparison over the reference genome <i>L. cremoris</i> MG1363 identified several unexpected mutations in the parental strain MG1614, likely selected during in-house propagation. In the Lcn972R clone, two previously identified mutations were mapped and confirmed. Additionally, another transposition event deregulating cellobiose uptake was identified along with three point mutations of unknown consequences for Lcn972 resistance. Two new independent evolution experiments exposing <i>L. cremoris</i> MG1614 to Lcn972 revealed transposition of <i>IS</i>981 into the <i>LLMG_RS12285</i> locus as the predominant mutation selected by Lcn972. This event occurs early during evolution and was found in 100% of the evolved clones, while other mutations were not selected. Therefore, activation of <i>LLMG_RS12285</i> coding for a putative anti-ECF (extra-cytoplasmic function) sigma factor is regarded as the main Lcn972 resistance factor in <i>L. cremoris</i> MG1614. |
| first_indexed | 2024-03-11T08:23:22Z |
| format | Article |
| id | doaj.art-a8620dc9f9b84ed4a026fa07671022d7 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-11T08:23:22Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-a8620dc9f9b84ed4a026fa07671022d72023-11-16T22:16:46ZengMDPI AGMicroorganisms2076-26072023-02-0111250110.3390/microorganisms11020501Resistance to the Bacteriocin Lcn972 Deciphered by Genome SequencingSusana Escobedo0Ana B. Campelo1Özgün C. O. Umu2María Jesús López-González3Ana Rodríguez4Dzung B. Diep5Beatriz Martínez6Instituto de Productos Lacteos de Asturias (IPLA), CSIC, 33300 Villaviciosa, SpainInstituto de Productos Lacteos de Asturias (IPLA), CSIC, 33300 Villaviciosa, SpainFaculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1430 Ås, NorwayInstituto de Productos Lacteos de Asturias (IPLA), CSIC, 33300 Villaviciosa, SpainInstituto de Productos Lacteos de Asturias (IPLA), CSIC, 33300 Villaviciosa, SpainFaculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1430 Ås, NorwayInstituto de Productos Lacteos de Asturias (IPLA), CSIC, 33300 Villaviciosa, SpainIn view of the current threat of antibiotic resistance, new antimicrobials with low risk of resistance development are demanded. Lcn972 is a lactococcal bacteriocin that inhibits septum formation by binding to the cell wall precursor lipid II in <i>Lactococcus</i>. It has a species-specific spectrum of activity, making Lcn972 an attractive template to develop or improve existing antibiotics. The aim of this work was to identify mutations present in the Lcn972-resistant clone <i>Lactococcus cremoris</i> D1-20, previously evolved from the sensitive strain <i>L. cremoris</i> MG1614. Whole-genome sequencing and comparison over the reference genome <i>L. cremoris</i> MG1363 identified several unexpected mutations in the parental strain MG1614, likely selected during in-house propagation. In the Lcn972R clone, two previously identified mutations were mapped and confirmed. Additionally, another transposition event deregulating cellobiose uptake was identified along with three point mutations of unknown consequences for Lcn972 resistance. Two new independent evolution experiments exposing <i>L. cremoris</i> MG1614 to Lcn972 revealed transposition of <i>IS</i>981 into the <i>LLMG_RS12285</i> locus as the predominant mutation selected by Lcn972. This event occurs early during evolution and was found in 100% of the evolved clones, while other mutations were not selected. Therefore, activation of <i>LLMG_RS12285</i> coding for a putative anti-ECF (extra-cytoplasmic function) sigma factor is regarded as the main Lcn972 resistance factor in <i>L. cremoris</i> MG1614.https://www.mdpi.com/2076-2607/11/2/501<i>Lactococcus</i>bacteriocinresistanceadaptive evolution |
| spellingShingle | Susana Escobedo Ana B. Campelo Özgün C. O. Umu María Jesús López-González Ana Rodríguez Dzung B. Diep Beatriz Martínez Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing Microorganisms <i>Lactococcus</i> bacteriocin resistance adaptive evolution |
| title | Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing |
| title_full | Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing |
| title_fullStr | Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing |
| title_full_unstemmed | Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing |
| title_short | Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing |
| title_sort | resistance to the bacteriocin lcn972 deciphered by genome sequencing |
| topic | <i>Lactococcus</i> bacteriocin resistance adaptive evolution |
| url | https://www.mdpi.com/2076-2607/11/2/501 |
| work_keys_str_mv | AT susanaescobedo resistancetothebacteriocinlcn972decipheredbygenomesequencing AT anabcampelo resistancetothebacteriocinlcn972decipheredbygenomesequencing AT ozguncoumu resistancetothebacteriocinlcn972decipheredbygenomesequencing AT mariajesuslopezgonzalez resistancetothebacteriocinlcn972decipheredbygenomesequencing AT anarodriguez resistancetothebacteriocinlcn972decipheredbygenomesequencing AT dzungbdiep resistancetothebacteriocinlcn972decipheredbygenomesequencing AT beatrizmartinez resistancetothebacteriocinlcn972decipheredbygenomesequencing |