Skip to main content

Advertisement

Springer Nature Link
Log in

Nutrient-Dependent Efficacy of the Antifungal Protein YvgO Correlates to Cellular Proliferation Rate in Candida albicans 3153A and Byssochlamys fulva H25

  • Published:
Probiotics and Antimicrobial Proteins Aims and scope Submit manuscript

Abstract

YvgO is a recently characterized antifungal protein isolated from Bacillus thuringiensis SF361 that exhibits a broad spectrum of activity and pH stability. Customized colorimetric metabolic assays based on standard broth microdilution techniques were used to determine the variable tolerance of Byssochlamys fulva H25 and Candida albicans 3153A to YvgO exposure under select matrix conditions impacting cellular proliferation. Normalization of the solution pH after antifungal challenge expanded the available pH range under consideration allowing for a comprehensive in vitro assessment of YvgO efficacy. Indicator susceptibility was examined across an array of elementary growth-modifying conditions, including media pH, incubation temperature, ionic strength, and carbohydrate supplementation. Under suboptimal temperature and pH conditions, the indicator growth rate reduced, and YvgO-mediated susceptibility was attenuated. While YvgO association but not efficacy was somewhat influenced by solution ionic strength, carbohydrate supplementation was shown to be the most influential susceptibility factor, particularly for C. albicans. Although the specific choice of carbohydrate/nutrient supplement dictated the extent of enhanced YvgO efficacy, d-glucose additionally improved the association between antifungal and target. Indeed, when exposed to YvgO under conditions that lead to increased cellular proliferation, both indicators displayed a stronger association and susceptibility to YvgO when compared to carbohydrate-deprived media or suboptimal incubation environments. With further study, YvgO may have the capacity to function as a prophylaxis for food safety and preservation, as well as a pharmaceutical agent against opportunistic fungal pathogens either independently or in combination with other established treatments applied to both livestock and human health concerns.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Explore related subjects

Discover the latest articles, books and news in related subjects, suggested using machine learning.

References

  1. Ben-Tal N, Honig B, Miller C, McLaughlin S (1997) Electrostatic binding of proteins to membranes. Theoretical predictions and experimental results with charybdotoxin and phospholipid vesicles. Biophys J 73(4):1717–1727

    Article  CAS  Google Scholar 

  2. Cantón E, Espinel-Ingroff A, Pemán J (2009) Trends in antifungal susceptibility testing using CLSI reference and commercial methods. Expert Rev Anti Infect Ther 7:107–119

    Article  Google Scholar 

  3. Chamilos G, Lewis RE, Kontoyiannis DP (2006) Inhibition of Candida parapsilosis mitochondrial respiratory pathways enhances susceptibility to caspofungin. Antimicrob Agents Chemother 50:744–747

    Article  CAS  Google Scholar 

  4. Clancy CJ, Nguyen MH (1997) Comparison of a photometric method with standardized methods of antifungal susceptibility testing of yeasts. J Clin Microbiol 35:2878–2882

    CAS  Google Scholar 

  5. Clinical and Laboratory Standards Institute (CLSI) (2008) Reference method for broth dilution antifungal susceptibility testing of filamentous fungi; approved standard, 2nd ed. CLSI document M38-A2. 28(16): 1–35

  6. Clinical and Laboratory Standards Institute (CLSI) (2008) Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard, 3rd ed. CLSI document M27-A3. 28(14):1–25

  7. Dent ME, Hubbold L, Radford H, Wilson AP (1996) Use of XTT for quantitating clonogenic growth in soft agar. Cytotechnology 18:219–225

    Article  Google Scholar 

  8. DeSilva B, Smith W, Weiner R, Kelly M, Smolec J, Lee B, Khan M, Tacey R, Hill H, Celniker A (2003) Recommendations for the bioanalytical method validation of ligand-binding assays to support pharmacokinetic assessments of macromolecules. Pharm Res 20:1885–1900

    Article  CAS  Google Scholar 

  9. Findlay JWA, Dillard RF (2007) Appropriate calibration curve fitting in ligand binding assays. AAPS J 9:E260–E267

    Article  Google Scholar 

  10. Finney DJ (1976) Radioligand assay. Biometrics 32:721–740

    Article  CAS  Google Scholar 

  11. Ghannoum MA, Rice LB (1999) Antifungal agents: mode of action, mechanisms of resistance, and correlation of these mechanisms with bacterial resistance. Clin Microbiol Rev 12:501–517

    CAS  Google Scholar 

  12. Hartvig RA, van de Weert M, Østergaard J, Jorgensen L, Jensen H (2011) Protein adsorption at charged surfaces: the role of electrostatic interactions and interfacial charge regulation. Langmuir 27(6):2634–2643

    Article  CAS  Google Scholar 

  13. Hawser SP, Norris H, Jessup CJ, Ghannoum MA (1998) Comparison of a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) colorimetric method with the standardized National Committee for Clinical Laboratory Standards method of testing clinical yeast isolates for susceptibility to antifungal agents. J Clin Microbiol 36:1450–1452

    CAS  Google Scholar 

  14. Kanafani ZA, Perfect JR (2008) Antimicrobial resistance: resistance to antifungal agents: mechanisms and clinical impact. Clin Infect Dis 46:120–128

    Article  Google Scholar 

  15. Kuhn DM, Balkis M, Chandra J, Mukherjee PK, Ghannoum MA (2003) Uses and limitations of the XTT assay in studies of Candida growth and metabolism. J Clin Microbiol 41:506–508

    Article  CAS  Google Scholar 

  16. Lee SM, Kim JM, Jeong J, Park YK, Bai GH, Lee EY, Lee MK, Chang CL (2007) Evaluation of the Broth Microdilution Method Using 2,3-Diphenyl-5-thienyl-(2)-tetrazolium Chloride for Rapidly Growing Mycobacteria Susceptibility Testing. J Korean Med Sci 22:784–790

    Article  CAS  Google Scholar 

  17. Lemaire K, Van de Velde S, Van Dijck P, Thevelein JM (2004) Glucose and sucrose act as agonist and mannose as antagonist ligands of the G protein-coupled receptor Gpr1 in the yeast Saccharomyces cerevisiae. Mol Cell 16:293–299

    Article  CAS  Google Scholar 

  18. Liu M, Seidel V, Katerere DR, Gray AI (2007) Colorimetric broth microdilution method for the antifungal screening of plant extracts against yeasts. Methods 42:325–329

    Article  CAS  Google Scholar 

  19. Maidan MM, Thevelein JM, Van Dijck P (2005) Carbon source induced yeast-to-hypha transition in Candida albicans is dependent on the presence of amino acids and on the G-protein-coupled receptor Gpr1. Biochem Soc Trans 33:291–293

    Article  CAS  Google Scholar 

  20. Manns DC, Churey JJ, Worobo RW (2012) Functional assignment of YvgO, a novel set of purified and chemically characterized proteinaceous antifungal variants produced by Bacillus thuringiensis SF361. Appl Environ Microb 78(8):2543–2552

    Article  CAS  Google Scholar 

  21. Meletiadis J, Meis JF, Mouton JW, Donnelly JP, Verweij PE (2000) Comparison of NCCLS and 3-(4, 5-dimethyl-2-Thiazyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) methods of in vitro susceptibility testing of filamentous fungi and development of a new simplified method. J Clin Microbiol 38:2949–2954

    CAS  Google Scholar 

  22. Meletiadis J, Mouton JW, Meis JF, Bouman BA, Donnelly JP, Verweij PE, Eurofung Network (2001) Colorimetric assay for antifungal susceptibility testing of Aspergillus species. J Clin Microbiol 39:3402–3408

    Article  CAS  Google Scholar 

  23. Paul KD, Shoemaker RH, Boyd MR (1988) The synthesis of XTT: a new tetrazolium reagent that is bioreducible to a water soluble formazan. J Heterocycl Chem 25:911–914

    Article  Google Scholar 

  24. Perea S, Patterson TF (2002) Antifungal Resistance in Pathogenic Fungi. Clin Infect Dis 35:1073–1080

    Article  Google Scholar 

  25. Rodaki A, Bohovych IM, Enjalbert B, Young T, Odds FC, Gow NAR, Brown AJP (2009) Glucose promotes stress resistance in the fungal pathogen Candida albicans. Mol Biol Cell 20:4845–4855

    Article  CAS  Google Scholar 

  26. Tellier R, Kajden M, Grigoriew GA, Campbell I (1992) Innovative endpoint determination system for antifungal susceptibility testing of yeasts. Antimicrob Agents Chemother 36:1619–1625

    Article  CAS  Google Scholar 

  27. Tournas VH, Heeres J, Burgess L (2006) Moulds and yeasts in fruit salads and fruit juices. Food Microbiol 23:684–688

    Article  CAS  Google Scholar 

  28. Towle HC (2005) Glucose as a regulator of eukaryotic gene transcription. Trends Endocrinol Metab 16:489–494

    Article  CAS  Google Scholar 

  29. Valeriote F, van Putten L (1975) Proliferation-dependent cytotoxicity of anticancer agents: a review. Cancer Res 35(10):2619–2630

    CAS  Google Scholar 

  30. Wang G, Manns DC, Guron GKP, Churey JJ, Worobo RW (2014) Large-scale purification, characterization, and spore outgrowth inhibitory effect of thurincin H, a bacteriocin produced by Bacillus thuringiensis SF361. Probiotics Antimicrob Proteins 6(2):105–113

    Article  CAS  Google Scholar 

Download references

Acknowledgments

Funding for this research was provided by the United States Department of Agriculture, National Integrated Food Safety Initiative (USDA-NIFSI) grant #2008-51110-0688 well as the National Science Foundation Graduate Research Fellowship Program. The authors would also like to thank the Cornell University Proteomics and Mass Spectrometry Core Facility and the Proteomics and Mass Spectrometry Facility at the Donald Danforth Plant Science Center for their technical assistance regarding initial YvgO purification.

Conflict of interest

The authors declare no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Food Science, New York State Agricultural Experiment Station, Cornell University, 630 West North Street, Geneva, NY, 14456, USA

    David C. Manns, John J. Churey & Randy W. Worobo

Authors
  1. David C. Manns
    View author publications

    Search author on:PubMed Google Scholar

  2. John J. Churey
    View author publications

    Search author on:PubMed Google Scholar

  3. Randy W. Worobo
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Randy W. Worobo.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Manns, D.C., Churey, J.J. & Worobo, R.W. Nutrient-Dependent Efficacy of the Antifungal Protein YvgO Correlates to Cellular Proliferation Rate in Candida albicans 3153A and Byssochlamys fulva H25. Probiotics & Antimicro. Prot. 6, 198–207 (2014). https://doi.org/10.1007/s12602-014-9167-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12602-014-9167-1

Keywords