Abstract
Peptides are important natural bioactive substances obtained from various sources such as microbes, plants and animals. The present study addressed the optimization and physicochemical characterization of the antibacterial peptide Marine Fungal Antibacterial Peptide 9 (MFAP9) produced by the marine fungus Aspergillus fumigatus BTMF9 and the in vitro evaluation of its antiproliferative, antioxidant and anti-inflammatory potential. Antiproliferative activity was evaluated in A549 lung carcinoma and normal L929 fibroblast cell lines using the MTT assay. Antioxidant activity was evaluated by DPPH assay and anti-inflammatory studies were performed by Cox2 inhibition assay on THP1 cell lines. The results showed that MFAP9 was cytotoxic against A549 cells with an IC50 value of 29 µg/mL and exhibited apoptotic properties after treatment with a concentration of 50 µg/mL. In addition, MFAP9 showed higher DPPH radical scavenging activity than standard antioxidants at the same concentration and significant anti-inflammatory activity. The rate of MFAP9 production was optimized by a time course experiment. Characterization revealed that the maximum active temperature of MFAP9 is 50 °C, that it is stable at basic pH, and that 1 mM concentrations of Ni2+, Cd2+, and Mg2+ promote peptide activity. It is hoped that MFAP9 could become a promising new therapeutic agent for various clinical applications by using advances in proteomics, bioinformatics and modification strategies.
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References
Agyei D, Danquah MK (2012) Rethinking food-derived bioactive peptides for antimicrobial and immunomodulatory activities. Trends Food Sci Technol 2:62–69. https://doi.org/10.1016/j.tifs.2011.08.010
Akindele AJ, Wani Z, Mahajan G et al (2014) Anticancer activity of Aristolochia ringens Vahl. (Aristolochiaceae). J Tradit Complement Med 5:35–41. https://doi.org/10.1016/j.jtcme.2014.05.001
Ambasht PK (2020) Use of group-specific reagents in active site functional group elucidation I: Cys, Ser, Tyr, and Trp residues. Springer, Singapore
Antolovich M, Prenzler PD, Patsalides E et al (2002) Methods for testing antioxidant activity. Analyst 127:183–198. https://doi.org/10.1039/B009171P
Arung ET, Shimizu K, Kondo R (2006) Inhibitory effect of artocarpanone from Artocarpus heterophyllus on melanin biosynthesis. Biol Pharm Bull 29:1966–1969. https://doi.org/10.1248/bpb.29.1966
Atalla MM, Zeinab H, Eman R et al (2008) Production of some biologically active secondary metabolites from marine-derived fungus Varicosporina ramulosa. Malays J Microbiol. https://doi.org/10.21161/MJM.01608
Atrih A, Rekhif N, Moir AJ et al (2001) Mode of action, purification and amino acid sequence of plantaricin C19, an anti-Listeria bacteriocin produced by Lactobacillus plantarum C19. Int J Food Microbiol 68:93–104. https://doi.org/10.1016/s0168-1605(01)00482-2
Attari F, Sepehri H, Delphi L, Goliaei B (2009) Apoptotic and necrotic effects of pectic acid on rat pituitary GH3/B6 tumor cells. Iran Biomed J 13:229–236
Bhugaloo-Vial P, Douliez J-P, Mollé D et al (1999) Delineation of key amino acid side chains and peptide domains for antimicrobial properties of divercin V41, a pediocin-like bacteriocin secreted by carnobacterium divergens V41. Appl Environ Microbiol 65:2895–2900
Birla SS, Gaikwad SC, Gade AK, Rai MK (2013) Rapid synthesis of silver nanoparticles from Fusarium oxysporum by optimizing physicocultural conditions. Sci World J 2013:796018. https://doi.org/10.1155/2013/796018
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Carniato F, Alberti D, Lapadula A et al (2019) Multifunctional Gd-based mesoporous silica nanotheranostic for anticancer drug delivery. J Mater Chem B 7:3143–3152. https://doi.org/10.1039/C9TB00375D
Čeponytė U, Paškevičiūtė M, Petrikaitė V (2018) Comparison of NSAIDs activity in COX-2 expressing and non-expressing 2D and 3D pancreatic cancer cell cultures. Cancer Manage Res 10:1543–1551. https://doi.org/10.2147/CMAR.S163747
Chakrabarti S, Jahandideh F, Wu J (2014) Food-derived bioactive peptides on inflammation and oxidative stress. Biomed Res Int 2014:e608979. https://doi.org/10.1155/2014/608979
Chanda S, Nagani K (2013) In vitro and in vivo Methods for Anticancer Activity Evaluation and Some Indian Medicinal Plants Possessing Anticancer Properties. An Overview. J Pharmacogn Phytochem 2:140
Chen Z, Ao J, Yang W et al (2013) Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment. Appl Microbiol Biotechnol 97:10381–10390. https://doi.org/10.1007/s00253-013-4800-6
Cheung RCF, Ng TB, Wong JH (2015) Marine peptides: bioactivities and applications. Mar Drugs 13:4006–4043. https://doi.org/10.3390/md13074006
Das S, Mishra B, Gill K et al (2011) Isolation and characterization of novel protein with anti-fungal and anti-inflammatory properties from Aloe vera leaf gel. Int J Biol Macromol 48:38–43. https://doi.org/10.1016/j.ijbiomac.2010.09.010
Diouf PN, Stevanovic T, Cloutier A (2009) Study on chemical composition, antioxidant and anti-inflammatory activities of hot water extract from Picea mariana bark and its proanthocyanidin-rich fractions. Food Chem 113:897–902
Eghtedari M, Jafari Porzani S, Nowruzi B (2021) Anticancer potential of natural peptides from terrestrial and marine environments: a review. Phytochem Lett 42:87–103. https://doi.org/10.1016/j.phytol.2021.02.008
Enan G, el-Essawy AA, Uyttendaele M, Debevere J (1996) Antibacterial activity of Lactobacillus plantarum UG1 isolated from dry sausage: characterization, production and bactericidal action of plantaricin UG1. Int J Food Microbiol 30:189–215. https://doi.org/10.1016/0168-1605(96)00947-6
Fu L, Zhou C, Yao S et al (2011) Plant lectins: targeting programmed cell death pathways as antitumor agents. Int J Biochem Cell Biol 43:1442–1449. https://doi.org/10.1016/j.biocel.2011.07.004
Halliwell B (1997) Antioxidants and human disease: a general introduction. Nutr Rev 55:S44-49. https://doi.org/10.1111/j.1753-4887.1997.tb06100.x
Hiroko Hassegawa R, Megumi Kasuya MC, Dantas Vanetti MC (2005) Growth and antibacterial activity of Lentinula edodes in liquid media supplemented with agricultural wastes. Electron J Biotechnol 8:94–99
Hohlfeld J, Fabel H, Hamm H (1997) The role of pulmonary surfactant in obstructive airways disease. Eur Respir J 10:482–491. https://doi.org/10.1183/09031936.97.10020482
Jaiganesh R, Sampath Kumar NS (2012) Marine bacterial sources of bioactive compounds. Adv Food Nutr Res 65:389–408. https://doi.org/10.1016/B978-0-12-416003-3.00025-1
Jain P, Pundir RK (2011) Effect of fermentation medium, pH and temperature variations on antibacterial soil fungal metabolite production. Int J Agric Technol 7:247–269
Joshi M, Patel H, Gupte S, Gupte A (2013) Nutrient improvement for simultaneous production of exopolysaccharide and mycelial biomass by submerged cultivation of Schizophyllum commune AGMJ-1 using statistical optimization. 3 Biotech 3:307–318. https://doi.org/10.1007/s13205-012-0103-3
Karimi E, Oskoueian E, Hendra R, Jaafar HZE (2010) Evaluation of Crocus sativus L. stigma phenolic and flavonoid compounds and its antioxidant activity. Molecules 15:6244–6256. https://doi.org/10.3390/molecules15096244
Kavitha Raja LC, Martin L, Bose et al (2020) Anti-proliferative and apoptotic effects of by-product (skin extract) from marine catfish Tachysurus dussumieri. Biocatal Agric Biotechnol 29:101816. https://doi.org/10.1016/j.bcab.2020.101816
Kawanishi T, Shiraishi T, Okano Y et al (2011) New detection systems of bacteria using highly selective media designed by SMART: selective medium-design algorithm restricted by two constraints. PLoS ONE 6:e16512. https://doi.org/10.1371/journal.pone.0016512
Khalil N, Ashour M, Singab AN, Salama O (2018) Bioassay guided fractionation and cytotoxic activity of Daucus carota var. boissieri. Future J Pharm Sci 4:14–17. https://doi.org/10.1016/j.fjps.2017.07.002
Kollakalnaduvil Raghavan RM, Ali Pannippara M, Kesav S et al (2021) MFAP9: characterization of an extracellular thermostable antibacterial peptide from marine fungus with biofilm eradication potential. J Pharm Biomed Anal 194:113808. https://doi.org/10.1016/j.jpba.2020.113808
Kumari M, Taritla S, Sharma A, Jayabaskaran C (2018) Antiproliferative and antioxidative bioactive compounds in extracts of marine-derived endophytic fungus Talaromyces purpureogenus. Front Microbiol 9:1777. https://doi.org/10.3389/fmicb.2018.01777
Lee YM, Kim MJ, Li H et al (2013) Marine-derived Aspergillus species as a source of bioactive secondary metabolites. Mar Biotechnol (NY) 15:499–519. https://doi.org/10.1007/s10126-013-9506-3
Liyana-Pathirana CM, Shahidi F (2005) Antioxidant activity of commercial soft and hard wheat (Triticum aestivum L.) as affected by gastric pH conditions. J Agric Food Chem 53:2433–2440. https://doi.org/10.1021/jf049320i
Luong HX, Thanh TT, Tran TH (2020) Antimicrobial peptides—advances in development of therapeutic applications. Life Sci 260:118407. https://doi.org/10.1016/j.lfs.2020.118407
Miao L, Qian P (2005) Antagonistic antimicrobial activity of marine fungi and bacteria isolated from marine biofilm and seawaters. Aquat Microbial Ecol. https://doi.org/10.3354/AME038231
Mogensen JE, Sehgal P, Otzen DE (2005) Activation, inhibition, and destabilization of Thermomyces lanuginosus lipase by detergents. Biochemistry 44:1719–1730. https://doi.org/10.1021/bi0479757
Ng TB (2005) Mushroom proteins related to host defense. Int J Med Mushroom. https://doi.org/10.1615/IntJMedMushr.v7.i12.210
Ottoni CA, Simões MF, Fernandes S et al (2017) Screening of filamentous fungi for antimicrobial silver nanoparticles synthesis. AMB Express 7:31. https://doi.org/10.1186/s13568-017-0332-2
Pannippara MA, Kesav S, Raghavan RMKN et al (2020) Characterization and potent application of Pleurotus floridanus trypsin inhibitor (PfTI). Nat Prod Sci 26:207–213. https://doi.org/10.20307/nps.2020.26.3.207
Petit KE, Mondeguer F, Roquebert MF et al (2004) Detection of griseofulvin in a marine strain of Penicillium waksmanii by ion trap mass spectrometry. J Microbiol Methods 58:59–65. https://doi.org/10.1016/j.mimet.2004.03.004
Raheema AS, Aswini C, Sathya M et al (2014) In-vitro anti-inflammatory screening of a poly herbal siddha medicine, “ASHWATHI CHOORANAM- In-Vitro Anti-Inflammatory screening of a poly herbal siddha medicine,“Aswathi Choornam". Int J Pharma Sci Res 5:4395
Rajasekar T, Balaji S, Kumaran S et al (2012) Isolation and characterization of Marine fungal metabolites against clinical pathogens. Asian Pac J Trop Dis 2:S387–S392. https://doi.org/10.1016/S2222-1808(12)60187-X
Robles-Fort A, García-Robles I, Fernando W et al (2021) Dual antimicrobial and antiproliferative activity of TcPaSK peptide derived from a Tribolium castaneum insect defensin. Microorganisms 9:222. https://doi.org/10.3390/microorganisms9020222
Silva FD, Rezende CA, Rossi DCP et al (2009) Structure and mode of action of microplusin, a copper II-chelating antimicrobial peptide from the cattle tick Rhipicephalus (Boophilus) microplus. J Biol Chem 284:34735–34746. https://doi.org/10.1074/jbc.M109.016410
Silva C, Correia-Branco A, Andrade N et al (2019) Selective pro-apoptotic and antimigratory effects of polyphenol complex catechin:lysine 1:2 in breast, pancreatic and colorectal cancer cell lines. Eur J Pharmacol 859:172533. https://doi.org/10.1016/j.ejphar.2019.172533
Skouri-Gargouri H, Jellouli-Chaker N, Gargouri A (2009) Factors affecting production and stability of the AcAFP antifungal peptide secreted by Aspergillus clavatus. Appl Microbiol Biotechnol. https://doi.org/10.1007/s00253-009-2279-y
Suarez-Jimenez G-M, Burgos-Hernandez A, Ezquerra-Brauer J-M (2012) Bioactive peptides and depsipeptides with anticancer potential: sources from marine animals. Mar Drugs 10:963–986. https://doi.org/10.3390/md10050963
Theis T, Wedde M, Meyer V, Stahl U (2003) The antifungal protein from Aspergillus giganteus causes membrane permeabilization. Antimicrob Agents Chemother 47:588–593. https://doi.org/10.1128/AAC.47.2.588-593.2003
Wang X, Yu H, Xing R, Li P (2017) Characterization, preparation, and purification of marine bioactive peptides. Biomed Res Int 2017:e9746720. https://doi.org/10.1155/2017/9746720
Wen C, Guo W, Chen X (2014) Purification and identification of a novel antifungal protein secreted by Penicillium citrinum from the Southwest Indian Ocean. J Microbiol Biotechnol 24:1337–1345. https://doi.org/10.4014/jmb.1405.05008
Xie J, Zhang R, Shang C-H, Guo Y (2009) Isolation and characterization of a bacteriocin produced by an isolated Bacillus subtilis LFB112 that exhibits antimicrobial activity against domestic animal pathogens. Afr J Biotechnol. https://doi.org/10.4314/AJB.V8I20.66016
Xu K, Gao H, Shu H-KG (2011) Celecoxib can induce vascular endothelial growth factor expression and tumor angiogenesis. Mol Cancer Ther 10:138–147. https://doi.org/10.1158/1535-7163.MCT-10-0415
Zan X, Cui F, Li Y et al (2015) Hericium erinaceus polysaccharide-protein HEG-5 inhibits SGC-7901 cell growth via cell cycle arrest and apoptosis. Int J Biol Macromol 76:242–253. https://doi.org/10.1016/j.ijbiomac.2015.01.060
Zhang H, Ji S, Guo R et al (2019) Hydrophobin HFBII-4 from Trichoderma asperellum induces antifungal resistance in poplar. Braz J Microbiol 50:603–612. https://doi.org/10.1007/s42770-019-00083-5
Zheng J, Zhu H, Hong K et al (2009) Novel cyclic hexapeptides from marine-derived fungus, Aspergillus sclerotiorum PT06-1. Org Lett 11:5262–5265. https://doi.org/10.1021/ol902197z
Acknowledgements
This work was supported by the Council of Scientific & Industrial Research (CSIR, New Delhi) under grant No. 10 − 2(5)/2007 (ii) – E.U.II in the form of fellowship to the first author.
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Raghavan, R.M.K., Pannippara, M.A., Kesav, S. et al. Production Optimization and In Vitro Evaluation of Anti-proliferative, Anti-oxidant and Anti-inflammatory Potential of the Antibacterial Peptide MFAP9. Int J Pept Res Ther 28, 139 (2022). https://doi.org/10.1007/s10989-022-10442-w
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DOI: https://doi.org/10.1007/s10989-022-10442-w