Abstract
With the emergence of antifungal resistance, systematic infections with Aspergillus are becoming the major cause of the clinical morbidity. The development of novel antifungal agents with high efficacy, low drug tolerance, and few side effects is urgent. In response to that need, we have identified NP20. Here we demonstrate clearly that NP20 has antifungal activity, capable of killing the spores of Aspergillus niger and Aspergillus fumigatus as well as causing direct damage to the surface, membrane, cytoplasm, organelle, and nucleus of the fungal spores. Interestingly, NP20 is active under temperature stress and a wide range of pH. Subsequently, MTT assay, assay for binding of NP20 to fungal cell wall components, membrane depolarization assay, confocal microscopy, ROS assay, DNA replication, and protein synthesis assay are performed to clarify the mechanisms underlying NP20 against Aspergillus. The results show that NP20 can bind with and pass through the fungal cell wall, and then interfere with the lipid membrane. Moreover, NP20 can induce intracellular ROS production, DNA fragmentation, and protein synthesis inhibition of the fungal cells. These together indicate that NP20 is a novel antifungal peptide, which has considerable potential for future development as novel peptide antibiotics against Aspergillus.
Similar content being viewed by others
References
Baddley JW, Stroud TP, Salzman D (2001) Invasive mold infections in allogeneic bone marrow transplant recipients. Clin Infect Dis 32:1319–1324
Bassetti M, Bouza E (2017) Invasive mould infections in the ICU setting: complexities and solutions. J Antimicrob Chemother 72(suppl_1):i39–i47
Brogden KA (2005) Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria?. Nat Rev Microbiol 3:238–250
Browne K, Chakraborty S, Chen R, Willcox MD, Black DS, Walsh WR, Kumar N (2020) A new era of antibiotics: the clinical potential of antimicrobial peptides. Int J Mol Sci 21:7047
Buda De Cesare G, Cristy SA, Garsin DA, Lorenz MC (2020) Antimicrobial peptides: a new frontier in antifungal therapy. mBio 11(6):e02123–20
Cadena J, Thompson GR 3rd, Patterson TF (2016) Invasive aspergillosis: current strategies for diagnosis and management. Infect Dis Clin North Am 30:125–142
Ciociola T, Giovati L, Conti S, Magliani W, Santinoli C, Polonelli L (2016) Natural and synthetic peptides with antifungal activity. Future Med Chem 8:1413–1433
Cui P, Dong Y, Li Z, Zhang Y, Zhang S (2016) Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum. Dev Comp Immunol 60:53–65
de Breij A, Riool M, Cordfunke RA, Malanovic N, de Boer L, Koning RI, Ravensbergen E, Franken M, van der Heijde T, Boekema BK, Kwakman PH (2018) The antimicrobial peptide SAAP-148 combats drug-resistant bacteria and biofilms. Sci Transl Med 10(423):eaan4044
De Brucker K, Cammue BP, Thevissen K (2011) Apoptosis-inducing antifungal peptides and proteins. Biochem Soc Trans 39:1527–1532
Duan H, Zhang X, Li Z, Yuan J, Shen F, Zhang S (2021) Synergistic effect and antibiofilm activity of an antimicrobial peptide with traditional antibiotics against multi-drug resistant bacteria. Microb Pathog 158:105056
Englund JA, Boeckh M, Kuypers J, Nichols WG, Hackman RC, Morrow RA, Fredricks DN, Corey L (2006) Brief communication: fatal human metapneumovirus infection in stem-cell transplant recipients. Ann Intern Med 144:344–349
Enoch DA, Ludlam HA, Brown NM (2006) Invasive fungal infections: a review of epidemiology and management options. J Med Microbiol 55:809–818
Erdem Büyükkiraz M, Kesmen Z (2021) Antimicrobial peptides (AMPs): a promising class of antimicrobial compounds. J Appl Microbiol
Feng LH, Li YQ, Wang ZS, Qi LQ, Mo HZ (2019) Antifungal actions of glycinin basic peptide against Aspergillus niger through the collaborative damage to cell membrane and mitochondria. Food Biophys 14:97–107
Gao Z, Qu B, Yao L, Ma Z, Cui P, Zhang S (2018) Identification and functional characterization of amphioxus Miple, ancestral type of vertebrate midkine/pleiotrophin homologues. Dev Comp Immunol 89:31–43
Girmenia C, Nucci M, Martino P (2001) Clinical significance of Aspergillus fungaemia in patients with haematological malignancies and invasive aspergillosis. Br J Haematol 114:93–98
Hegedüs N, Marx F (2013) Antifungal proteins: more than antimicrobials? Fungal Biol Rev 26:132–145
Latgé JP (1999) Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev 12:310–350
Latgé JP, Chamilos G (2019) Aspergillus fumigatus and Aspergillosis in 2019. Clin Microbiol Rev 33:e00140-e218
Lee H, Hwang JS, Lee J, Kim JI, Lee DG (2015) Scolopendin 2, a cationic antimicrobial peptide from centipede, and its membrane-active mechanism. Biochim Biophys Acta 1848:634–642
Li C, Jia X, Bian Y, Qi D, Wu J (2021) Different susceptibility of spores and hyphae of Trichophyton rubrum to methylene blue mediated photodynamic treatment in vitro. Mycoses 64:48–54
Maribel PJ, Gustavo V, Roberto O, Maria Monica CO, Keiko S (2003) Effect of chitosan and temperature on spore germination of Aspergillus niger. Macromol Biosci 3:582–586
Ning HQ, Li YQ, Tian QW, Wang ZS, Mo HZ (2019) The apoptosis of Staphylococcus aureus induced by glycinin basic peptide through ROS oxidative stress response. LWT Food Sci Technol 99:62–68
Nivoix Y, Ledoux MP, Herbrecht R (2020) Antifungal therapy: new and evolving therapies. Semin Respir Crit Care Med 41:158–174
Prasad R, Shah AH, Rawal MK (2016) Antifungals: mechanism of action and drug resistancE. Adv Exp Med Biol 892:327–349
Perfect JR (2017) The antifungal pipeline: a reality check. Nat Rev Drug Discov 16:603–616
Perrone G, Gallo A (2017) Aspergillus species and their associated mycotoxins. Methods Mol Biol 1542:33–49
Rhodes JC, Jensen HE, Nilius AM, Chitambar CR, Farmer SG, Washburn RG, Steele PE, Amlung TW (1992) Aspergillus and aspergillosis. J Med Vet Mycol 30:51–57
Scorzoni L, de Paula E, Silva AC, Marcos CM, Assato PA, de Melo WC, de Oliveira HC, Costa-Orlandi CB, Mendes-Giannini MJ, Fusco-Almeida AM (2017) Antifungal therapy: new advances in the understanding and treatment of mycosis. Front Microbiol 8:36
Sorrelle N, Dominguez ATA, Brekken RA (2017) From top to bottom: midkine and pleiotrophin as emerging players in immune regulation. J Leukoc Biol 102:277–286
Sugui JA, Kwon-Chung KJ, Juvvadi PR, Latgé JP, Steinbach WJ (2014) Aspergillus fumigatus and related species. Cold Spring Harb Perspect Med 5:a019786
Sun Q, Li J, Sun Y, Chen Q, Zhang L, Le T (2020) The antifungal effects of cinnamaldehyde against Aspergillus niger and its application in bread preservation. Food Chem 317:126405
Svensson SL, Pasupuleti M, Walse B, Malmsten M, Mörgelin M, Sjögren C, Olin AI, Collin M, Schmidtchen A, Palmer R, Egesten A (2010) Midkine and pleiotrophin have bactericidal properties: preserved antibacterial activity in a family of heparin-binding growth factors during evolution. J Biol Chem 285:16105–16115
Taccone FS, Van den Abeele AM, Bulpa P, Misset B, Meersseman W, Cardoso T, Paiva JA, Blasco-Navalpotro M, De Laere E, Dimopoulos G, Rello J (2015) Epidemiology of invasive aspergillosis in critically ill patients: clinical presentation, underlying conditions, and outcomes. Crit Care 19:7
Vandewoude K, Blot S, Benoit D, Depuydt P, Vogelaers D, Colardyn F (2004) Invasive aspergillosis in critically ill patients: analysis of risk factors for acquisition and mortality. Acta Clin Belg 59:251–257
van der Weerden NL, Bleackley MR, Anderson MA (2013) Properties and mechanisms of action of naturally occurring antifungal peptides. Cell Mol Life Sci 70:3545–3570
Wang K, Yan J, Dang W, Xie J, Yan B, Yan W, Sun M, Zhang B, Ma M, Zhao Y, Jia F, Zhu R, Chen W, Wang R (2014) Dual antifungal properties of cationic antimicrobial peptides polybia-MPI: membrane integrity disruption and inhibition of biofilm formation. Peptides 56:22–29
Wang Y, Cui P, Zhang Y, Yang Q, Zhang S (2018) Augmentation of the antibacterial activities of Pt5-derived antimicrobial peptides (AMPs) by amino acid substitutions: design of novel AMPs against MDR bacteria. Fish Shellfish Immunol 77:100–111
Zhang QX, Zhang Y, Shan HH, Tong YH, Chen XJ, Liu FQ (2017) Isolation and identification of antifungal peptides from Bacillus amyloliquefaciens W10. Environ Sci Pollut Res 24:25000–25009
Acknowledgements
The authors acknowledge the substantive input from all members of the Laboratory for Evolution & Development.
Funding
This work was supported by the Ministry of Science and Technology (MOST) of China (grant numbers 2018YFD0900505) and the Marine S&T Fund of Shandong province for Pilot National Laboratory for Marine Science and Technology (Qingdao) (grant numbers 2018SDKJ0302-1).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics Approval
The authors confirm that the ethical policies of the journal, as noted on the journal’s author guidelines page, have been adhered to. No ethical approval was required as the research in this article related to microorganisms.
Conflict of Interest
The authors declare no competing interests. The authors alone are responsible for the content and the writing of the paper.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Li, Z., Shen, F., Song, L. et al. Antifungal Activity of NP20 Derived from Amphioxus Midkine/Pleiotrophin Homolog Against Aspergillus niger and Aspergillus fumigatus. Mar Biotechnol 24, 614–625 (2022). https://doi.org/10.1007/s10126-022-10131-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10126-022-10131-1