Analysis of the cell wall binding domain in bacteriocin-like lysin LysL from Lactococcus lactis LAC460.
Lactococcus lactis
Cell wall binding domain
Green fluorescent protein
Prophage lysin
Journal
Archives of microbiology
ISSN: 1432-072X
Titre abrégé: Arch Microbiol
Pays: Germany
ID NLM: 0410427
Informations de publication
Date de publication:
02 Jul 2024
02 Jul 2024
Historique:
received:
27
03
2024
accepted:
21
06
2024
revised:
19
06
2024
medline:
2
7
2024
pubmed:
2
7
2024
entrez:
2
7
2024
Statut:
epublish
Résumé
Wild-type Lactococcus lactis strain LAC460 secretes prophage-encoded bacteriocin-like lysin LysL, which kills some Lactococcus strains, but has no lytic effect on the producer. LysL carries two N-terminal enzymatic active domains (EAD), and an unknown C-terminus without homology to known domains. This study aimed to determine whether the C-terminus of LysL carries a cell wall binding domain (CBD) for target specificity of LysL. The C-terminal putative CBD region of LysL was fused with His-tagged green fluorescent protein (HGFPuv). The HGFPuv_CBDlysL gene fusion was ligated into the pASG-IBA4 vector, and introduced into Escherichia coli. The fusion protein was produced and purified with affinity chromatography. To analyse the binding of HGFPuv_CBDLysL to Lactococcus cells, the protein was mixed with LysL-sensitive and LysL-resistant strains, including the LysL-producer LAC460, and the fluorescence of the cells was analysed. As seen in fluorescence microscope, HGFPuv_CBDLysL decorated the cell surface of LysL-sensitive L. cremoris MG1614 with green fluorescence, whereas the resistant L. lactis strains LM0230 and LAC460 remained unfluorescent. The fluorescence plate reader confirmed the microscopy results detecting fluorescence only from four tested LysL-sensitive strains but not from 11 tested LysL-resistant strains. Specific binding of HGFPuv_CBDLysL onto the LysL-sensitive cells but not onto the LysL-resistant strains indicates that the C-terminus of LysL contains specific CBD. In conclusion, this report presents experimental evidence of the presence of a CBD in a lactococcal phage lysin. Moreover, the inability of HGFPuv_CBDLysL to bind to the LysL producer LAC460 may partly explain the host's resistance to its own prophage lysin.
Identifiants
pubmed: 38954047
doi: 10.1007/s00203-024-04066-5
pii: 10.1007/s00203-024-04066-5
doi:
Substances chimiques
Bacteriocins
0
Green Fluorescent Proteins
147336-22-9
Recombinant Fusion Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
336Subventions
Organisme : Niemi-säätiö
ID : 20210081
Organisme : The Finnish Cultural Foundation
ID : 27.02.2022
Informations de copyright
© 2024. The Author(s).
Références
Abdelrahman F, Easwaran M, Daramola OI, Ragab S, Lynch S, Oduselu TJ et al (2021) Phage-encoded endolysins. Antibiotics 10:124. https://doi.org/10.3390/antibiotics10020124
doi: 10.3390/antibiotics10020124
pubmed: 33525684
pmcid: 7912344
Ainsworth S, Sadovskaya I, Vinogradov E, Courtin P, Guerardel Y, Mahony J et al (2014) Differences in lactococcal cell wall polysaccharide structure are major determining factors in bacteriophage sensitivity. Mbio 5:e00880-e1814. https://doi.org/10.1128/mBio.00880-14
doi: 10.1128/mBio.00880-14
pubmed: 24803515
pmcid: 4010823
Bobay LM, Touchon M, Rocha EPC (2014) Pervasive domestication of defective prophages by bacteria. PNAS 111:12127–12132. https://doi.org/10.1073/pnas.1405336111
doi: 10.1073/pnas.1405336111
pubmed: 25092302
pmcid: 4143005
Broendum SS, Ashley MB, Sheena M (2018) Catalytic diversity and cell wall binding repeats in the phage-encoded endolysins. Mol Microbiol 110:879–896. https://doi.org/10.1111/mmi.14134
doi: 10.1111/mmi.14134
pubmed: 30230642
Chandran C, Tham HY, Abdul RR, Lim SHE, Yusoff K, Song AA (2022) Lactococcus lactis secreting phage lysins as a potential antimicrobial against multi-drug resistant Staphylococcus aureus. PeerJ 10:e12648. https://doi.org/10.7717/peerj.12648
doi: 10.7717/peerj.12648
pubmed: 35251775
pmcid: 8896023
Chopin A, Chopin MC, Moillo-Batt A, Langella P (1984) Two plasmid-determined restriction and modification systems in Streptococcus lactis. Plasmid 11:260–263. https://doi.org/10.1016/0147-619x(84)90033-7
doi: 10.1016/0147-619x(84)90033-7
pubmed: 6087394
Chopin A, Deveau H, Ehrlich SD, Moineau S, Chopin M (2007) KSY1, a lactococcal phage with a T7-like transcription. Virol 365:1–9. https://doi.org/10.1016/j.virol.2007.03.044
doi: 10.1016/j.virol.2007.03.044
Colombo A (2016) Reformulation and quality evaluation by microbiological analyses of idli, a fermented Indian products. MSc thesis, Università Degli Studi di Milano, Italy, and University of Helsinki, Finland
Desvaux M, Candela T, Serror P (2018) Surfaceome and proteosurfaceome in parietal monoderm bacteria: focus on protein cell-surface display. Front Microbiol 9:100. https://doi.org/10.3389/fmicb.2018.00100
doi: 10.3389/fmicb.2018.00100
pubmed: 29491848
pmcid: 5817068
Dorosky RJ, Lola SL, Brown HA, Schreier JE, Dreher-Lesnick SM, Stibitz S (2023) Characterization of lactobacilli phage endolysins and their functional domains-potential live biotherapeutic testing reagents. J Viruses 15:1986. https://doi.org/10.3390/v15101986
doi: 10.3390/v15101986
Edwards U, Rogall T, Blöcker H, Emde M, Böttger EC (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16s ribosomal RNA. Nucleic Acids Res 17(19):7843–7853. https://doi.org/10.1093/nar/17.19.7843
doi: 10.1093/nar/17.19.7843
pubmed: 2798131
pmcid: 334891
Efstathiou JD, McKay LL (1977) Inorganic salts resistance associated with a lactose-fermenting plasmid in Streptococcus lactis. J Bacteriol 130:257–265. https://doi.org/10.1128/jb.130.1.257-265.1977
doi: 10.1128/jb.130.1.257-265.1977
pubmed: 404284
pmcid: 235201
Eugster MR, Loessner MJ (2012) Wall teichoic acids restrict access of bacteriophage endolysin Ply118, Ply511, and Plyp40 cell wall binding domains to the Listeria monocytogenes peptidoglycan. J Bacteriol 194:6498–6506. https://doi.org/10.1128/JB.00808-12
doi: 10.1128/JB.00808-12
pubmed: 23002226
pmcid: 3497465
Eugster MR, Haug MC, Huwiler SGB, Loessner MJ (2011) The cell wall binding domain of Listeria bacteriophage endolysin PlyP35 recognizes terminal GlcNAc residues in cell wall teichoic acid. Mol Microbiol 81:1419–1432. https://doi.org/10.1111/j.1365-2958.2011.07774.x
doi: 10.1111/j.1365-2958.2011.07774.x
pubmed: 21790805
Gasson MJ (1983) Plasmid complements of Streptococcus lactis NCDO 712 and other lactic streptococci after protoplast-induced curing. J Bacteriol 154:1–9. https://doi.org/10.1128/jb.154.1.1-9.1983
doi: 10.1128/jb.154.1.1-9.1983
pubmed: 6403500
pmcid: 217423
Goh S, Ong PF, Song KP, Riley TV, Chang BJ (2007) The complete genome sequence of Clostridium difficile phage phiC2 and comparisons to phiCD119 and inducible prophages of CD630. Microbiol (reading) 153:676–685. https://doi.org/10.1099/mic.0.2006/002436-0
doi: 10.1099/mic.0.2006/002436-0
Hakovirta J, Reunanen J, Saris PEJ (2006) Bioassay for nisin in milk, processed cheese, salad dressings, canned tomatoes, and liquid egg products. AEM 72:1001–1005. https://doi.org/10.1128/AEM.72.2.1001-1005.2006
doi: 10.1128/AEM.72.2.1001-1005.2006
Hanahan D (1985) Techniques for transformation of Escherichia coli. In: Glover DM (ed) DNA cloning: a practical approach. IRL Press, Virginia, pp 109–135
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. https://doi.org/10.1038/227680a0
doi: 10.1038/227680a0
pubmed: 5432063
Loessner MJ (2005) Bacteriophage endolysins–current state of research and applications. COMICR 8:480–487. https://doi.org/10.1016/j.mib.2005.06.002
doi: 10.1016/j.mib.2005.06.002
Loessner MJ, Kramer K, Ebel F, Scherer S (2002) C-terminal domains of Listeria monocytogenes bacteriophage murein hydrolases determine specific recognition and high-affinity binding to bacterial cell wall carbohydrates. Mol Microbiol 44:335–349. https://doi.org/10.1046/j.1365-2958.2002.02889.x
doi: 10.1046/j.1365-2958.2002.02889.x
pubmed: 11972774
Lortal S, Chapot-Chartier MP (2005) Role, mechanisms and control of lactic acid bacteria lysis in cheese. Int Dairy J 15:857–871. https://doi.org/10.1016/j.idairyj.2004.08.024
doi: 10.1016/j.idairyj.2004.08.024
Mahony J, Kot W, Murphy J, Ainsworth S, Neve H, Hansen LHK et al (2013) Investigation of the relationship between lactococcal host cell wall polysaccharide genotype and 936 phage receptor binding protein phylogeny. AEM 79:4385–4392. https://doi.org/10.1128/AEM.00653-13
doi: 10.1128/AEM.00653-13
Oechslin F, Zhu X, Dion MB, Shi R, Moineau S (2022) Phage endolysins are adapted to specific hosts and are evolutionarily dynamic. PLoS Biol 20:e3001740. https://doi.org/10.1371/journal.pbio.3001740
doi: 10.1371/journal.pbio.3001740
pubmed: 35913996
pmcid: 9371310
Oliveira H, Melo LD, Santos SB, Nóbrega FL, Ferreira EC, Cerca N et al (2013) Molecular aspects and comparative genomics of bacteriophage endolysins. J Virol 87:4558–4570. https://doi.org/10.1128/JVI.03277-12
doi: 10.1128/JVI.03277-12
pubmed: 23408602
pmcid: 3624390
Oliveira H, São-José C, Azeredo J (2018) Phage-derived peptidoglycan degrading enzymes: challenges and future prospects for in vivo therapy. J Viruses 10:292. https://doi.org/10.3390/v10060292
doi: 10.3390/v10060292
Plavec TV, Štrukelj B, Berlec A (2019) Screening for new surface anchoring domains for Lactococcus lactis. Front Microbiol 10:1879. https://doi.org/10.3389/fmicb.2019.01879
doi: 10.3389/fmicb.2019.01879
pubmed: 31456787
pmcid: 6700490
Raya-Tonetti F, Müller M, Sacur J, Kitazawa H, Villena J, Vizoso-Pinto MG (2021) Novel LysM motifs for antigen display on lactobacilli for mucosal immunization. Sci Rep 11:21691. https://doi.org/10.1038/s41598-021-01087-8
doi: 10.1038/s41598-021-01087-8
pubmed: 34737363
pmcid: 8568972
Schmelcher M, Tchang VS, Loessner MJ (2011) Domain shuffling and module engineering of Listeria phage endolysins for enhanced lytic activity and binding affinity. Microb Biotechnol 4:651–662. https://doi.org/10.1111/j.1751-7915.2011.00263.x
doi: 10.1111/j.1751-7915.2011.00263.x
pubmed: 21535426
pmcid: 3819014
Shin H, Lee JH, Yoon H, Kang DH, Ryu S (2014) Genomic investigation of lysogen formation and host lysis systems of the Salmonella temperate bacteriophage SPN9CC. AEM 80:374–384. https://doi.org/10.1128/AEM.02279-13
doi: 10.1128/AEM.02279-13
Sorokina D (2015) Enkasiini-bakteriosiinin tuotto Escherichia coli-bakteerissa. MSc thesis, University of Helsinki. Finland
Steen MT, Chung YJ, Hansen JN (1991) Characterization of the nisin gene as part of a polycistronic operon in the chromosome of Lactococcus lactis ATCC 11454. Appl Environ Microbiol 57:1181–1188. https://doi.org/10.1128/aem.57.4.1181-1188.1991
doi: 10.1128/aem.57.4.1181-1188.1991
pubmed: 1905517
pmcid: 182865
Sui B, Qi X, Wang X, Ren H, Liu W, Zhang C (2021) Characterization of a novel bacteriophage swi2 harboring two lysins can naturally lyse Escherichia coli. Front Microbiol 12:670799. https://doi.org/10.3389/fmicb.2021.670799
doi: 10.3389/fmicb.2021.670799
pubmed: 34113331
pmcid: 8185280
Summer EJ, Berry J, Tran TA, Niu L, Struck DK, Young R (2007) Rz/Rz1 lysis gene equivalents in phages of Gram-negative hosts. J Mol Biol 373:1098–1112. https://doi.org/10.1016/j.jmb.2007.08.045
doi: 10.1016/j.jmb.2007.08.045
pubmed: 17900620
Takác M, Bläsi U (2005) Phage P68 virion-associated protein 17 displays activity against clinical isolates of Staphylococcus aureus. Antimicrob Agents Chemother 49:2934–2940. https://doi.org/10.1128/AAC.49.7.2934-2940
doi: 10.1128/AAC.49.7.2934-2940
pubmed: 15980371
pmcid: 1168661
Takala TM, Mokhtari S, Ahonen SL, Wan X, Saris PEJ (2023) Wild-type Lactococcus lactis producing bacteriocin-like prophage lysins. Front Microbiol 14:1219723. https://doi.org/10.3389/fmicb.2023.1219723
doi: 10.3389/fmicb.2023.1219723
pubmed: 37520360
pmcid: 10377672
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. PNSA 76:4350–4354. https://doi.org/10.1073/PNAS.76.9.4350
doi: 10.1073/PNAS.76.9.4350
Wan X, Takala TM, Qiao M, Saris PEJ (2021) Complete genome sequence of nisin-producing Lactococcus lactis subsp. lactis N8. Microbiol Resour Announc 10:e01147-e1220. https://doi.org/10.1128/MRA.01147-20
doi: 10.1128/MRA.01147-20
pubmed: 33414301
pmcid: 8407701
Zabarovsky ER, Winberg G (1990) High efficiency electroporation of ligated DNA into bacteria. Nucleic Acids Res 18:5912. https://doi.org/10.1093/nar/18.19.5912
doi: 10.1093/nar/18.19.5912
pubmed: 2216800
pmcid: 332358
Zahirović A, Plavec TV, Berlec A (2022) Dual functionalized Lactococcus lactis shows tumor antigen targeting and cytokine binding in vitro. Front Bioeng Biotechnol 10:822823. https://doi.org/10.3389/fbioe.2022.822823
doi: 10.3389/fbioe.2022.822823
pubmed: 35155394
pmcid: 8826564