Optimization of growth parameters for enhancing antifungal secondary metabolites of Talaromyces islandicus VSGF1 against drug resistant Candida spp.

Abstract

The rapid increase in resistance to antifungal drugs has been raised largely in Candida species due to continue and over utilization of available frontline drugs. To overcome this rising health care issue, it’s necessary to identify a new and improved antifungal drug without causing any side effects than the existing drugs. In view of this, the present investigation was carried out in findings of the novel antifungal drug against pathogenic Candida species. The fungus Talaromyces islandicus VSGF1 was screened for the production of antifungal secondary metabolites through optimization of growth parameters under submerged fermentation by changing one variable at a time. The highest fungal biomass (1.9g/100ml of culture broth) with significant antifungal activity (15 mm zone of inhibition) was achieved in potato dextrose broth supplemented with 2% dextrose at pH6 and 26 OC temperature at stationary condition. A comparative antifungal activity was performed on frontline drugs of amphotericin B, fluconazole, ketoconazole, and nystatin along with extracellular and intracellular crude extracts of T. islandicus against eight Candida strains. Among the frontline drugs, only ketoconazole exhibited antifungal activity against tested Candida spp. and resistance has been observed by amphotericin B, fluconazole, and nystatin. Interestingly, both extracts of T. islandicus were found to possess significant and a broad spectrum of inhibitory activity against all tested Candida spp. This is the first report on this species from the genus Talaromyces that was used for the optimization of process parameters and contribution to the potential producers for antifungal secondary metabolites.

References

1. Frisvad JC. Taxonomy, chemodiversity and chemoconsistency of Aspergillus, Penicillium and Talaromyces species. Front. Microbiol. 2015; 5(1):1-7.
2. Marino T, Nishimoto M, Masuda A, Fujita S, Nishikiori T, Saito S. NK374200, a novel insecticidal agent from Talaromyces, found by physico-chemical screening. J. Antibiot. 1995; 48(12):1509-10.
3. Thrane U, Rasmussen KB, Petersen B, Rasmussen S, Sicheritz-Ponten, Mortensen UH. Genome sequence of Talaromyces atroroseus, which produces red colorants for the food industry. Genome Announc. 2017; 5(9):1736-16.
4. Hong LL, Xiao ML, Hui L, Ling HM, Bin G W. Two new diphenylketones and a new xanthone from Talaromyces islandicus EN-501, an endophytic fungus derived from the marine red algae Laurencia okamurai. Mar. Drugs. 2016; 14:1-8.
5. Sanchez JF, Somoza AD, Keller NP, Wang CCC. Advances in Aspergillus secondary metabolite research in the post-genomic era. Nat. Prod. Rep. 2012; 29(3):351–71.
6. Brakhage AA. Regulation of fungal secondary metabolism. Nat. Rev. Microbiol. 2013; 11:21–32.
7. Scherlach K, Graupner K, Hertweck C. Molecular bacteria-fungi interactions: effects on environment, food and medicine. Annu. Rev. Microbiol. 2013; 67:375–97.
8. Takahashi JA, Teles APC, Bracarense AAP, Gomes DC. Classical and epigenetic approaches to metabolite diversification in filamentous fungi. Phytochem. Rev. 2013; 12:773–89.
9. Bertrand S, Bohni N, Schnee S, Schumpp O, Gindro K, Wolfender JL. Metabolite induction via microorganism co-culture: a potential way to enhance chemical diversity for drug discovery. Biotechnol. Adv. 2014; 32(6):1180–204.
10. Marmann A, Aly A H, Lin W, Wang B, Proksch P. Co-cultivation – a powerful emerging tool for enhancing the chemical diversity of microorganisms. Mar. Drugs. 2014; 12(2):1043–62.
11. Berkow EL, Lockhart SR. Fluconazole resistance in Candida species: a current perspective. Infect. Drug. Resist. 2017; 10:237-45.
12. Gao J, Wang H, Li Z, Wong AHH, Wang YZ, Guo Y, Lin X, Zeng G, Liu H, Wang Y, Wang J. Candida albicans gains azole resistance by altering sphingolipid composition. Nat. Commun. 2018; 9(1):4495.
13. Deorukhkar SC, Roushani S. Fluconazole resistance in Candida species: ten years experience at a rural tertiary care teaching hospital in India. J. Infect. Dis. and Pathog. 2017; 1:102.
14. Sachafhauser T, Wibberg D, Ruckert C, Winkler A, Flor L, van Pee KH, Fewer DP, Sivonen K, Jahn L, Ludwig-Muller J, Caradec T, Jacques P, Hujibers MM, van Berkel WJ, Weber T, Wohlleben W, Kalinowski J. Draft genome sequence of Talaromyces islandicus (“Penicillium islandicus”) WF-38-12, a neglected mold with significant biotechnological potential. J. Biotechnol. 2015; 211:101-2.
15. Mathan S, Subramanian V, Nagamony S. Optimization and antimicrobial metabolite production from endophytic fungi Aspergillus KC 582297. Euro. J. Exp. Biol. 2013; 3:138-44.
16. Asnaashari M, Ghanbary MAT, Tazick Z. Optimization of penicillin G production by Penicillium chrysogenum. Ann. Biol. Res. 2012; 3(12):5434-40.
17. Cao C, Li R, Wan Z, Liu W, Wang X, Qiao J, Wang D, Bulmer G, Calderone R. The effects of temperature, pH, and salinity on the growth and dimorphism of Penicillium marneffei. Med. Mycol. 2007; 45(5):401-7.
18. Ritchie F, Bain RA, McQuilken MP. Effects of nutrient status, temperature and pH on mycelia growth, sclerotial production and germination of Rhizoctonia solani from potato. J. Plant. Pathol. 2009; 91(3):589-96.
19. Compaore H, Sawadogo HL, Guira F, Ware L Y, Samandoulougou S, Savadogo A, Dianou D, Traore AS. Optimization of antimicrobial compounds production by Aspergillus fumigates isolated from maize in Ouagadougou, Burkina Faso. Current Res. Microbiol. Biotechnol. 2016; 4(4):903-11.
20. Al-Shaibani ABA, Al-Shakarchi FI, Ameen RS. Extraction and characterization of antibacterial compound from Aspergillus niger. AI-Nahrain J. Sci. 2013; 16(4):167-74.
21. Magaldi S, Mata-Essayag S, Capriles CH, Perez C, Colella MT, Olaizola C, Ontiveros Y. Well diffusion for antifungal susceptibility testing. Int. J. Infect. Dis. 2004; 8(1):39-45.
22. Valgas C, de Souza SM, Smania EFA, Smania A. Screening methods to determine antibacterial activity of natural products. Braz. J. Microbiol. 2007; 38(2):369-80.
23. Noura NEE, Haroun SA, Oweis EA, Sherief AA. Identification of newly isolated Talaromyces pinophilus and statistical optimization of β-Glucosidase production under solid state fermentation. Preparative Biochem. Biotechnol. 2015; 45(7): 712-29.
24. Gowthaman MK, Krishna C, Moo-Young M. Fungal solid state fermentation-an overview. Appl. Mycol. Biotechnol. 2001; 1:305-52.
25. Barrios-Gonzalez J, Tarrago-castellanos MR. Solid-State fermentation: special physiology of fungi. In: Merillon JM., Ramawat K. (eds) Fungal metabolites. References series in phytochemistry. Springer, Cham. 2017; 319-347.
26. Yamazaki M, Okuyama E. Isolation and structure of oxaphenalenone dimmers from Talaromyces bacillosporus. Chem. Pharm. Bull. 1980; 28(12): 3649-55.
27. Bhattacharyya PN, Jha DK. Optimization of cultural conditions affecting growth and improved bioactive metabolite production by a subsurface Aspergillus strain TSF 146. Int. J. Appl. Biol. Pharm. Technol. 2011; 2(4):133-45.
28. Verma SK, Lal M, Debnath M. Optimization of process parameters for production of antimicrobial metabolites by endophytic fungus Aspergillus sp. CPR5 isolated from Calotropis procera root. Asian J. Pharma. Sci. Clin. Res. 2017; 10(4):225-30.
29. Mustafa U, Kaur G. Effects of carbon and nitrogen source and ratio on the germination, growth and sporulation characteristics of Metarhizium anisopliae and Beauveria bassiana isolates. Afr. J. Agric. Res. 2009; 3(10):922-30.
30. Gao L, Sun MH, Liu XZ, Che YS. Effects of carbon concentration and carbon to nitrogen ratio on the growth and sporulation of several bio-control fungi. Mycol. Res. 2007; 111(1):87-92.
31. Hassan SAA, Bakhiet SEA. Optimization of antibacterial compounds production by Aspergillus fumigates isolated from Sudanese indigenous soil. Int. Biol. Biomed. J. 2017; 3(4):204-8.
32. Gogoi DK, Deka Boruah HP, Saikia R, Bora TC. Optimization of process parameters for improved production of bioactive metabolite by a novel endophytic fungus Fusarium sp. DF2 isolated from Taxus wallichiana. World J. Microbiol. Biotechnol. 2008; 24(1):79-87.
33. Jain P, Pundir RK. Effect of fermentation medium, pH and temperature variations on antibacterial soil fungal metabolite production. Int. J. Agric. Technol. 2011; 7(2):247-69.
34. Palanichamy P, Krishnamoorthy G, Kannan S, Marudhamuthu M. Bioactive potential of secondary metabolites derived from medicinal plant endophytes. Egypt. J. Basic and Appl. Sci. 2018; 5(4):303–12.
35. Meier CL, Rapp J, Bowers RM, Silman M, Fierer N. Fungal growth on a common wood substrate across a tropical elevation gradient: Temperature sensitivity, community composition, and potential for above-ground decomposition. Soil Biol. Biochem. 2010; 42(7):1083–90.
36. Sharma A, Sharma M, Chandra S. Influence of temperature and relative humidity on growth and sporulation of some common dermatophytes. Indian J. Fund. Appl. Life Sci. 2012; 2(4):1-6.
37. Vijayakumar R, Panneerselvam K, Muthukumar C, Thajuddin N, Paneerselvam A, Saravanamuthu R. Optimization of antimicrobial production by a marine actinomycetes Streptomyces sfghaniensis VPTS3-1 isolated from palk strait, east coast of India. Indian J. Microbiol. 2012; 52(2): 230-39.
38. Pfaller MA, Diekema DJ, Gibbs DL, Newell VA, Ellis D, Tullio V, Rodloff A, Fu W, Ling TA. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2007: a 10.5- year analysis of susceptibilities of Candida species to fluconazole and voriconazole as determined by CLSI standardized disk diffusion. J. Clin. Microbiol. 2010; 48(4):1366–77.
39. Nolte FS, Parkinson T, Falconer DJ, Dix S, Williams J, Gilmore C, Eller R, Wingard JR. Isolation and characterization of fluconazole- and amphotericin B–resistant Candida albicans from blood of two patients with leukemia. Antimicrob. Agents Chemother. 1997; 41(1):196–9.
40. Anaissie EJ, Hachem R, Legrand C, Legenne P, Nelson P, Bodey GP. Lack of activity of amphotericin B in systemic murine fusarial infection. J. Infect. Dis. 1992; 165(6):1155–7.
41. Ellis D. Amphotericin B: spectrum and resistance. J. Antimicrob. Chemother. 2002; 49(1):7–10.
42. Powderly WG, Kobayashi GS, Herzig GP, Medoff G. Amphotericin B–resistant yeast infection in severely immunocompromised patients. Am. J. Med. 1988; 84(5):826–32.
43. Ahmadi A, Mahmoudi S, Rezaie S, Hashemi SJ, Dannaoui E, Badali H, Ghaffari M, Aala F, Izadi A, Maleki A, Meis JF, Khodavaisy S. In vitro synergy of echinocandins with triazoles against fluconazole-resistant Candida parapsilosis complex isolates. J. Glob. Antimicrob. Resist. 2020; 21:313-334.
44. Ahangarkani F, Khodavaisy S, Mahmoudi S, Shokohi T, Rezai MS, Fakhim H, Dannaoui E, Faraji S, Choedhary A, Meis JF, Badali H. Indifferent effect of nonsteroidal anti-inflammatory drugs (NSAIDs) combined with fluconazole against multidrug-resistant Candida auris. Curr. Med. Mycol. 2019; 5(3):26–30.
45. Giang V, Tran H, Nguyen A, Nguyen Y, Nguyen T. Identification, antimicrobial activities and cytotoxicity of Talaromyces purpurogenus T013. J. Chem. Pharm. Res. 2017; 9(12): 174-9.
46. Pretsch A, Nagl M, Schwendinger K, Kreiseder B, Wiederstein M, Pretsh D, Genov M, Hollaus R, Zinssmeister D, Debbab A, Hundsberger H, Eger A, Proksch P, Wiesner C. Antimicrobial and anti-inflammatory activities of endophytic fungi Talaromyces wortmannii extracts against acne-inducing bacteria. PLoS ONE. 2014; 9(6).
47. Miao F, Yang R, Chen D, Wang Y, Qin B, Yang X, Zhou L. Isolation, identification and antimicrobial activities of two secondary metabolites of Talaromyces verruculosus. Molecules. 2012; 17: 14091-8.
48. Suzuki S, Hosoe T, Nozawa K, Kawai K, Yaguchi T, Udagawa S. Antifungal substances against pathogenic fungi, Talaroconvolutins, from Talaromyces convolutes. J. Nat. Prod. 2000; 63(6):768-72.
49. Suzuki S, Nozawa K, Nakajima S, Udagawa S, Kawai K. Isolation and structure of antibacterial binaphtho-a-pyrones, Talaroderxines A and B, from Talaromyces derxii. Chem. Pharm. Bull. 1992; 40(5):1116-9.
50. Ayer WA, Racok JS. The metabolites of Talaromyces flavus: Part 2. Biological activity and biosynthetic studies. Can. J. Chem. 1990; 68(11):2095-101.
51. Komai S, Hosoe T, Tabashi T, Nozawa K, Okada K, Takaki GMC, Chikamori M, Yaguchi T, Fukushima K, Miyaji M, Kawai K. A new funicone derivative isolated from Talaromyces flavus IFM52668. Mycotoxin. 2004; 54(1):15-9.
52. Arai M, Tomoda H, Okuda T, Wang H, Tabata N, Masuma R, Yamaguchi Y, Omura S. Funicone related compounds, potentiators of antifungal miconazole activity produced by Talaromyces flavus FKI-0076. J. Antibiot. 2002; 55(2):172-80.