Abstract
Plantaricin NC8, a two-peptide non-lantibiotic class IIb bacteriocin composed of PLNC8α and PLNC8β and derived from Lactobacillus plantarum ZJ316, has been shown to be highly potent against a range of bacteria and fungi. In this study, we assessed the antimicrobial mechanism of plantaricin NC8 against the most sensitive bacterial strain, Micrococcus luteus CGMCC 1.193. The results showed that plantaricin NC8 induced membrane permeabilization and caused cell membrane disruption to M. luteus CGMCC 1.193 cells, as evidenced by electrolyte efflux, loss of proton motive force, and ATP depletion within a few minutes of plantaricin NC8 treatment. Furthermore, scanning and transmission electron microscopy showed that plantaricin NC8 had a drastic impact on the structure and integrity of M. luteus CGMCC 1.193 cells. In addition, we found that either PLNC8α or PLNC8β alone exhibited membrane permeabilization activity, but that PLNC8β had higher permeabilization activity, and their individual effects were not as strong as that of the combined compounds as plantaricin NC8. Finally, we showed that lipid II is not the specific target of plantaricin NC8 against M. luteus CGMCC 1.193. Our study reveals the antimicrobial mechanism of plantaricin NC8 against M. luteus CGMCC 1.193.
Similar content being viewed by others
References
Abee T, Klaenhammer TR, Letellier L (1994) Kinetic studies of the action of lactacin F, a bacteriocin produced by Lactobacillus johnsonii that forms poration complexes in the cytoplasmic membrane. Appl Environ Microbiol 60:1006–1013
Breukink E (2003-01-15) Method for preparing lipid II and use of the lipid II thus obtained. EP:EP1275731. http://www.freepatentsonline.com/EP1275731A1.html
Breukink E, de Kruijff B (2006) Lipid II as a target for antibiotics. Nat Rev Drug Discov 5:321–332. https://doi.org/10.1038/nrd2004
Breukink E, Wiedemann I, Van KC, Kuipers OP, Sahl HG, De KB (1999) Use of the cell wall precursor lipid II by a pore-forming peptide antibiotic. Science 286:2361–2364. https://doi.org/10.1126/science.286.5448.2361
Brötz H, Bierbaum G, Leopold K, Reynolds PE, Sahl HG (1998) The lantibiotic mersacidin inhibits peptidoglycan synthesis by targeting lipid II. Antimicrob Agents Chemother 42:154–160
Castellano P, Raya R, Vignolo G (2003) Mode of action of lactocin 705, a two-component bacteriocin from Lactobacillus casei CRL705. Int J Food Microbiol 85:35–43. https://doi.org/10.1016/S0168-1605(02)00479-8
Coelho MLV, Duarte AFD, Bastos MDD (2017) Bacterial labionin-containing peptides and sactibiotics: unusual types of antimicrobial peptides with potential use in clinical settings (a review). Curr Top Med Chem 17:1177–1198. https://doi.org/10.2174/1568026616666160930144809
Cuozzo SA, Castellano P, Sesma FJ, Vignolo GM, Raya RR (2003) Differential roles of the two-component peptides of lactocin 705 in antimicrobial activity. Curr Microbiol 46:180–183. https://doi.org/10.1007/s00284-002-3844-0
Diep DB, Skaugen M, Salehian Z, Holo H, Nes IF (2007) Common mechanisms of target cell recognition and immunity for class II bacteriocins. PNAS 104:2384–2389. https://doi.org/10.1073/pnas.0608775104
Essig A, Hofmann D, Münch D, Gayathri S, Künzler M, Kallio PT, Sahl HG, Wider G, Schneider T, Aebi M (2014) Copsin, a novel peptide-based fungal antibiotic interfering with the peptidoglycan synthesis. J Biol Chem 289:34953–34964. https://doi.org/10.1074/jbc.M114.599878
Gabrielsen C, Brede DA, Hernandez PE, Nes IF, Diep DB (2012) The maltose ABC transporter in Lactococcus lactis facilitates high-level sensitivity to the circular bacteriocin Garvicin ML. Antimicrob Agents Chemother 56:2908–2915. https://doi.org/10.1128/Aac.00314-12
Jiang H, Li P, Gu Q (2016) Heterologous expression and purification of plantaricin NC8, a two-peptide bacteriocin against Salmonella, spp. from Lactobacillus plantarum ZJ316. Protein Expr Purif 127:28–34. https://doi.org/10.1016/j.pep.2016.06.013
Jiang H, Zou J, Cheng H, Fang J, Huang G (2017) Purification, characterization, and mode of action of pentocin JL-1, a novel bacteriocin isolated from Lactobacillus pentosus, against drug-resistant Staphylococcus aureus. Biomed Res Int 2017:7657190. https://doi.org/10.1155/2017/7657190
Kalchayanand N, Dunne P, Sikes A, Ray B (2004) Viability loss and morphology change of foodborne pathogens following exposure to hydrostatic pressures in the presence and absence of bacteriocins. Int J Food Microbiol 91:91–98. https://doi.org/10.1016/S0168-1605(03)00324-6
Khalaf H, Nakka SS, Sandén C, Svärd A, Hultenby K, Scherbak N, Aili D, Bengtsson T (2016) Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis. BMC Microbiol 16:188. https://doi.org/10.1186/s12866-016-0810-8
Kjos M, Oppegard C, Diep DB, Nes IF, Veening JW, Nissen-Meyer J, Kristensen T (2014) Sensitivity to the two-peptide bacteriocin lactococcin G is dependent on UppP, an enzyme involved in cell-wall synthesis. Mol Microbiol 92(6):1177–1187. https://doi.org/10.1111/mmi.12632
Li X, Gu Q, Lou X, Zhang X, Song D, Shen L, Zhao Y (2013) Complete genome sequence of the probiotic Lactobacillus plantarum strain ZJ316. Genome Announc 1:e00094–e00013. https://doi.org/10.1128/genomeA.00094-13
Ling LL, Schneider T, Peoples AJ, Spoering AL, Engels I, Conlon BP, Mueller A, Schaberle TF, Hughes DE, Epstein S, Jones M, Lazarides L, Steadman VA, Cohen DR, Felix CR, Fetterman KA, Millett WP, Nitti AG, Zullo AM, Chen C, Lewis K (2015) A new antibiotic kills pathogens without detectable resistance. Nature 517(7535):455. https://doi.org/10.1038/nature14098
Maldonado A, Ruizbarba JL, Jiménezdíaz R (2003) Purification and genetic characterization of plantaricin NC8, a novel coculture-inducible two-peptide bacteriocin from Lactobacillus plantarum NC8. Appl Environ Microbiol 69:383–389. https://doi.org/10.1128/AEM.69.1.383-389.2003
Marciset O, Jeronimus-Stratingh MC, Mollet B, Poolman B (1997) Thermophilin 13, a nontypical antilisterial poration complex bacteriocin, that functions without a receptor. J Biol Chem 272:14277–14284. https://doi.org/10.1074/jbc.272.22.14277
Moll G, Hildeng-Hauge H, Nissen-Meyer J, Nes IF, Konings WN, Driessen AJM (1998) Mechanistic properties of the two-component, bacteriocin lactococcin G. J Bacteriol 180:96–99
Moll GN, den AE V, Hauge HH, Nissen-Meyer J, Nes IF, Konings WN, Driessen ALM (1999) Complementary and overlapping selectivity of the two-peptide bacteriocins plantaricin EF and JK. J Bacteriol 181:4848–4852
Nes IF, Holo H (2000) Class II antimicrobial peptides from lactic acid bacteria. Biopolymers 55:50–61. https://doi.org/10.1002/1097-0282(2000)55:1<50::AID-BIP50>3.0.CO;2-3
Nicolas P (2009) Multifunctional host defense peptides: intracellular-targeting antimicrobial peptides. FEBS J 276:6483–6496. https://doi.org/10.1111/j.1742-4658.2009.07359.x
Nissen-Meyer J, Oppegard C, Rogne P, Haugen HS, Kristiansen PE (2010) Structure and mode-of-action of the two-peptide (class-IIb) bacteriocins. Probiotics Antimicro Proteins 2:52–60. https://doi.org/10.1007/s12602-009-9021-z
Pattanayaiying R, H-Kittikun A, Cutter CN (2014) Effect of lauric arginate, nisin Z, and a combination against several food-related bacteria. Int J Food Microbiol 188:135–146. https://doi.org/10.1016/j.ijfoodmicro.2014.07.013
Roces C, Courtin P, Kulakauskas S, Rodríguez A, Chapot-Chartier MP, Martínez B (2012) Isolation of Lactococcus lactis mutants simultaneously resistant to the cell wall-active bacteriocin Lcn972, lysozyme, nisin, and bacteriophage c2. Appl Environ Microbiol 78:4157–4163. https://doi.org/10.1128/AEM.00795-12
Schneider T, Kruse T, Wimmer R, Wiedemann I, Sass V, Pag U, Jansen A, Nielsen AK, Mygind PH, Raventós DS, Neve S, Ravn B, Bonvin AM, De Maria L, Andersen AS, Gammelgaard LK, Sahl HG, Kristensen HH (2010) Plectasin, a fungal defensin, targets the bacterial cell wall precursor Lipid II. Science 328:1168–1172. https://doi.org/10.1126/science.1185723
Uzelac G, Kojic M, Lozo J, Aleksandrzak-Piekarczyk T, Gabrielsen C, Kristensen T, Nes IF, Diep DB, Topisirovic L (2013) A Zn-dependent metallopeptidase is responsible for sensitivity to LsbB, a class II leaderless bacteriocin of Lactococcus lactis subsp lactis BGMN1–5. J Bacteriol 195(24):5614–5621. https://doi.org/10.1128/Jb.00859-13
Valenzuela JF, Pinuer LA, Cancino AG, Yáñez RB (2015) Metabolic fluxes in lactic acid bacteria-a review. Food Biotechnol 29:185–217
van Kan EJ, Demel RA, Breukink E, Van der Bent A, de Kruijff B (2002) Clavanin permeabilizes target membranes via two distinctly different pH-dependent mechanisms. Biochemistry 41:7529–7539. https://doi.org/10.1021/bi012162t
Varney KM, Bonvin AMJJ, Pazgier M, Malin J, Yu W, Ateh E, Oashi T, Lu W, Huang J, Buin MD, Bryant J, Breukink E, Mackerell AD, de Leeuw EPH (2013) Turning defense into offense: defensin mimetics as novel antibiotics targeting lipid II. PLoS Pathog 9:e1003732. https://doi.org/10.1371/journal.ppat.1003732
Wiedemann I, Breukink E, Van KC, Kuipers OP, Bierbaum G, De KB, Sahl HG (2001) Specific binding of nisin to the peptidoglycan precursor lipid II combines pore formation and inhibition of cell wall biosynthesis for potent antibiotic activity. J Biol Chem 276:1772–1779. https://doi.org/10.1074/jbc.M006770200
Wiedemann I, Böttiger T, Bonelli RR, Wiese A, Hagge SO, Gutsmann T, Seydel U, Deegan L, Hill C, Ross P, Sahl HG (2006a) The mode of action of the lantibiotic lacticin 3147--a complex mechanism involving specific interaction of two peptides and the cell wall precursor lipid II. Mol Microbiol 61:285–296. https://doi.org/10.1111/j.1365-2958.2006.05223.x
Wiedemann I, Böttiger T, Bonelli RR, Schneider T, Sahl HG, Martínez B (2006b) Lipid II-based antimicrobial activity of the lantibiotic plantaricin C. Appl Environ Microbiol 72:2809–2814. https://doi.org/10.1128/AEM.72.4.2809-2814.2006
Zhang X, Wang Y, Liu L, Wei Y, Shang N, Zhang X, Li P (2016) Two-peptide bacteriocins PlnEF causes cell membrane damage to Lactobacillus plantarum. Biochim Biophys Acta 1858:274–280. https://doi.org/10.1016/j.bbamem.2015.11.018
Zhou K, Zhou W, Li P, Dai Y (2008) Mode of action of pentocin 31-1: an antilisteria bacteriocin produced by Lactobacillus pentosus from Chinese traditional ham. Food Control 19:817–822. https://doi.org/10.1016/j.foodcont.2007.08.008
Zhu X, Zhao Y, Sun Y, Gu Q (2014) Purification and characterisation of plantaricin ZJ008, a novel bacteriocin against Staphylococcus spp. from Lactobacillus plantarum ZJ008. Food Chem 165:216–223. https://doi.org/10.1016/j.foodchem.2014.05.034
Funding
This project was funded by the National Science Foundation of China (No. 31601449), Natural Science Foundation of Zhejiang Province (No. LY16C200002, No. LQ18C200004), International Science & Technology Cooperation Program of China (No. 2013DFA32330), the Major Science and Technology Projects of Zhejiang Province (2015C02039, 2015C02022), and the Food Science and Engineering—the most important discipline of Zhejiang Province (2017SIAR202).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Rights and permissions
About this article
Cite this article
Jiang, H., Tang, X., Zhou, Q. et al. Plantaricin NC8 from Lactobacillus plantarum causes cell membrane disruption to Micrococcus luteus without targeting lipid II. Appl Microbiol Biotechnol 102, 7465–7473 (2018). https://doi.org/10.1007/s00253-018-9182-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-018-9182-3