EVALUATION OF DIFFERENT PROTEIN PRECIPITATES OF A WILD MUSHROOM, GANODERMA LUCIDUM FOR ANTIBACTERIAL ACTIVITIES AGAINST HUMAN PATHOGENIC BACTERIA
DOI:
https://doi.org/10.22159/ajpcr.2019.v12i7.33714Keywords:
Mushroom,, Acid extract,, Trichloroacetic acid,, Minimum inhibitory volume,, Hemolytic assayAbstract
Objective: The objective of the present study was to isolate different antibacterial protein precipitates from Ganoderma lucidum against human pathogenic bacteria and to evaluate suitable precipitating agent.
Methods: The acid extract was prepared from the aqueous solution of the test mushroom. From separate aliquots of acid extract, antibacterial proteins were precipitated using five different concentrations (10–50%) of ammonium sulfate solutions, 10% trichloroacetic acid (TCA), 80% ethanol and methanol – Chloroform mixture (2:1 ratio). Protein quantification was performed in each stage of purifications. The as-prepared protein precipitates were subjected for antibacterial and hemolytic assays for identification of the active protein precipitate, which in turn was also checked for minimum inhibitory volume (MIV) for all test organisms.
Results: The quantity of each protein precipitated by different protein precipitating agents from the acid extract of the test mushroom was found in the range of 2.3–4.8 mg/g wet.wt. Although all the precipitates showed different levels of antibacterial capacities, 10% TCA precipitate was considered as active protein as it yielded the maximum amount of protein (4.8 mg/g.wet.wt) as well as it exhibited burly bactericidal activities at lower volumes of protein solutions subjected (6.3 and 3.2 μl) on all bacterial strains tested with less hemolytic effects.
Conclusion: The protein precipitated by 10% TCA from the acid extract of the test mushroom could be developed as a drug candidate for treating infectious diseases caused by pathogenic microbes in human.
Downloads
References
Harbarth S, Harris AD, Carmeli Y, Samore MH. Parallel analysis of individual and aggregated data on antibiotic exposure and resistance in gram negative bacilli. Clin Infect Dis 2001;33:1462-8.
Segal-Maurer S, Urban C, Rahal JJ Jr. Current perspectives on multidrug-resistant bacteria. Epidemiology and control. Infect Dis Clin North Am 1996;10:939-57.
Hamid IS, Eltayeb IM. In vitro susceptibility of Shigella flexneri and Shigella dysenteriae to the ethanolic extracts of Trachysperum ammi and Peganum harmala. Int J Pharm Pharm Sci 2019;11:103-6.
Dahham SS, Hew CS, Jaafar I, Gam LH. The protein profiling of Asian giant toad skin secretions and their antimicrobial activity. Int J Pharm Pharm Sci 2016;8:88-95.
Sundaramoorthy M, Saravanan TS. Antibacterial effects of goat and chicken heart tissues against human pathogenic bacteria. Indian J Exp Biol 2010;48:407-14.
Daher KA, Selsted ME, Lehrer RI. Direct inactivation of viruses by human granulocyte defensins. J Virol 1986;60:1068-74.
Diamond G, Zasloff M, Eck H, Brasseur M, Maloy WL, Bevins CL, et al. Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: Peptide isolation and cloning of a cDNA. Proc Natl Acad Sci U S A 1991;88:3952-6.
Selsted ME, Tang YQ, Morris WL, McGuire PA, Novotny MJ, Smith W, et al. Purification, primary structures, and antibacterial activities of beta-defensins, a new family of antimicrobial peptides from bovine neutrophils. J Biol Chem 1993;268:6641-8.
Harder J, Bartels J, Christophers E, Schröder JM. A peptide antibiotic from human skin. Nature 1997;387:861.
Rinaldi AC. Antimicrobial peptides from amphibian skin: An expanding scenario. Curr Opin Chem Biol 2002;6:799-804.
Sundaramoorthy M, Saravanan TS. A natural antibacterial peptide from goat heart tissues against S. typhi and P. aeruginosa. World J Zool 2011;6:227-34.
Sundaramoorthy M, Saravanan TS. Purification of an antibactierial peptide from chicken heart tissues by RP- HPLC against Pseudomonas aeruginosa. Int J Sci Nat 2011;2:310-6.
Sundaramoorthy M, Gomathi S, Kannaki S, Saravanan TS. Antibacterial and haemolytic activities of tissue proteins of the major carp, Labeo Rohita (Hamilton). Int J Sci Nat 2011;2:671-6.
Zaiou YT, Gallo R. Some bioactive properties of wild and commercial mushroom species. J Food Health Sci 2002;3:161-9.
Bals R. Wild and commercial mushrooms as source of nutrients and nutraceuticals. Food Chem Toxicol 1998;46:2742-7.
Houshdar Tehrani MH, Fakhrehoseini E, Kamali Nejad M, Mehregan H, Hakemi-Vala M. Search for proteins in the liquid extract of edible mushroom, Agaricus bisporus, and studying their antibacterial effects. Iran J Pharm Res 2012;11:145-50.
Pouchert P, Fons F, Raiors S. Biological and pharmacological activity of higher fungi 20-year retrospective analysis. Mycologie 2006;27:311 33.
Öztürk M, Duru ME, Kivrak S, Mercan-Doğan N, Türkoglu A, Özler MA, et al. In vitro antioxidant, anticholinesterase and antimicrobial activity studies on three Agaricus species with fatty acid compositions and iron contents: A comparative study on the three most edible mushrooms. Food Chem Toxicol 2011;49:1353-60.
Zheng S, Liu Q, Zhang G, Wang H, Ng TB. Purification and characterization of an antibacterial protein from dried fruiting bodies of the wild mushroom Clitocybe sinopica. Acta Biochim Pol 2010;57:43 8.
Hearst M, Nelson D, McCollum G, Linda M, Ballard B, Moore S, et al. Antimicrobial properties of protein extracts from wild mushroom fungi and native plant species against hospital pathogens. J Pharmacogn Phytother 2010;2:103-7.
Wasser SP. Reishi or Lingzhi (Ganoderma lucidum) In: Encyclopedia of Dietary Supplements. New York: Marcel Decker; 2005. p. 603-20.
Lindequist U, Niedermeyer TH, Jülich WD. The pharmacological potential of mushrooms. Evid Based Complement Alternat Med 2005;2:285-99.
Sa-Ard P, Sarnthima R, Khammuang S, Kanchanarach W. Antioxidant, antibacterial and DNA protective activities of protein extracts from Ganoderma lucidum. J Food Sci Technol 2015;52:2966-73.
Matutte B, Storey KB, Knoop FC, Conlon JM. Induction of synthesis of an antimicrobial peptide in the skin of the freeze-tolerant frog, Rana sylvatica, in response to environmental stimuli. FEBS Lett 2000;483:135-8.
Sundaramoorthy M, Deepa K, Saravanan TS. Chemical preparation of bioactive protein from cow heart tissues: An evaluation. Int J Adv Res Biotech 2012;1:18-21.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265-75.
Sarker SD, Nahar L, Kumarasamy Y. Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods 2007;42:321-4.
Zhu WL, Hahm KS, Shin SY. Cathelicidin-derived trp/Pro-rich antimicrobial peptides with lysine peptoid residue (Nlys): Therapeutic index and plausible mode of action. J Pept Sci 2007;13:529-35.
Jagadesh LK, Krishnan V, Shenbhagaraman R Kaviyarasan V. Comparative study on the antioxidant, anticancer and antimicrobial property of Agaricus bisporus (J.E Lange) Imbach before and after boiling. Afr J Biotechnol 2009;8:654-61.
Ramesh Ch, Pattar MG. Antimicrobial properties, antioxidant activity and bioactive compounds from six wild edible mushrooms of Western Ghats of Karnataka, India. Pharmacognosy Res 2010;2:107-12.
Lacadena J, Martínez del Pozo A, Gasset M, Patiño B, Campos-Olivas R, Vázquez C, et al. Characterization of the antifungal protein secreted by the mould Aspergillus giganteus. Arch Biochem Biophys 1995;324:273- 81.
Lee DG, Shin SY, Maeng CY, Jin ZZ, Kim KL, Hahm KS, et al. Isolation and characterization of a novel antifungal peptide from Aspergillus niger. Biochem Biophys Res Commun 1999;263:646-51.
Weiler F, Schmitt MJ. Zygocin, a secreted antifungal toxin of the yeast Zygosaccharomyces bailii, and its effect on sensitive fungal cells. FEMS Yeast Res 2003;3:69-76.
Guo Y, Wang H, Ng TB. Isolation of trichogin, an antifungal protein from fresh fruiting bodies of the edible mushroom Tricholoma giganteum. Peptides 2005;26:575-80.
Xue Q, Ding Y, Shang C, Jiang C, Zhao M. Functional expression of LZ-8, a fungal immunomodulatory protein from Ganoderma lucidium in Pichia pastoris. J Gen Appl Microbiol 2008;54:393-8.
Singha SS, Soltnian H, Razaenin A, Janpoor S. Antibactrial activity of crude extract and fractions from Iranian wild-grown and cultivation. Biomed Res 2017;27:56-9.
Lam SK, Ng TB. First simultaneous isolation of a ribosome inactivating protein and an antifungal protein from a mushroom (Lyophyllum shimeji) together with evidence for synergism of their antifungal effects. Arch Biochem Biophys 2001;393:271-80.
Kavitha R, Damodharan N. Ultrasonic assisted extraction and antibacterial activities of protein recovered from white Button Mushroom Agaricus bisporus. Int J PharmTech Res 2015;8:991-5.
Coban EP, Isman B, Biyik HH. Effects of Protein bound polysaccharide against some microorganism. Turk Mikrobial Cem Derg 2008;38:150 5.
Xiao Y, Dai H, Bommineni YR, Soulages JL, Gong YX, Prakash O, et al. Structure-activity relationships of fowlicidin-1, a cathelicidin antimicrobial peptide in chicken. FEBS J 2006;273:2581-93.
Falla TJ, Karunaratne DN, Hancock RE. Mode of action of the antimicrobial peptide indolicidin. J Biol Chem 1996;271:19298-303.
Yang ST, Shin SY, Kim Y, Comformation dependent antimicrobial peptide. Biochem Biophys Res Commun 2002;296:1044-105.
Published
How to Cite
Issue
Section
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.
