Determination of the concentrations of possible endogenous photosensitizers in <i>Candida auris</i> via two-dimensional fluorescence spectroscopy

Anna-Maria Gierke; Robin Haag; Martin Hessling; Anna-Maria Gierke; Robin Haag; Martin Hessling

doi:10.3934/biophy.2025007

AIMS Biophysics

2025, Volume 12, Issue 1: 101-120. doi: 10.3934/biophy.2025007

Previous Article Next Article

Research article

Determination of the concentrations of possible endogenous photosensitizers in Candida auris via two-dimensional fluorescence spectroscopy

Institute of Medical Engineering and Mechatronics, Ulm University of Applied Sciences, Albert-Einstein-Allee 55, D-89081 Ulm, Germany

Received: 01 December 2024 Revised: 27 January 2025 Accepted: 17 February 2025 Published: 17 March 2025

Candida auris is an emerging pathogenic yeast. In order to investigate possible light-based disinfection systems or therapeutic approaches, this study examined the formation of possible photosensitizers under different growth conditions. The yeast was cultivated under aerobic conditions in YEPG medium and minimal medium with 10% NaCl. 2D fluorescence spectroscopy (15 excitation wavelengths from 320 to 450 nm) at different time points was performed to calculate the concentrations and proportions of dry mass of various fluorophores. Comparing the different growth conditions, higher concentrations of porphyrins were observed with minimal medium + 10% NaCl than with the YEPG medium. When determining the concentrations of different fluorophores over time, different dynamics could be observed for both aerobic growth conditions. For both growth conditions, NADH, followed by thiamine and lumichrome, exhibited the highest concentrations; coproporphyrin III showed the lowest.
- photosensitizer,
- fluorophores,
- fluorescence measurements,
- concentration,
- ROS,
- flavin,
- porphyrin
Citation: Anna-Maria Gierke, Robin Haag, Martin Hessling. Determination of the concentrations of possible endogenous photosensitizers in Candida auris via two-dimensional fluorescence spectroscopy[J]. AIMS Biophysics, 2025, 12(1): 101-120. doi: 10.3934/biophy.2025007

Related Papers:

Abstract

Candida auris is an emerging pathogenic yeast. In order to investigate possible light-based disinfection systems or therapeutic approaches, this study examined the formation of possible photosensitizers under different growth conditions. The yeast was cultivated under aerobic conditions in YEPG medium and minimal medium with 10% NaCl. 2D fluorescence spectroscopy (15 excitation wavelengths from 320 to 450 nm) at different time points was performed to calculate the concentrations and proportions of dry mass of various fluorophores. Comparing the different growth conditions, higher concentrations of porphyrins were observed with minimal medium + 10% NaCl than with the YEPG medium. When determining the concentrations of different fluorophores over time, different dynamics could be observed for both aerobic growth conditions. For both growth conditions, NADH, followed by thiamine and lumichrome, exhibited the highest concentrations; coproporphyrin III showed the lowest.

Conflict of interest

The authors declare no conflict of interest.

Author contributions

Conceptualization: A.-M.G. and M.H.; methodology, A.-M.G.; software, A.-M.G.; validation, A.-M.G., R.H. and M.H.; formal analysis, A.-M.G.; investigation, A.-M.G. and R.H.; resources, M.H.; data curation, A.-M.G.; writing—original draft preparation, A.-M.G.; writing—review and editing, A.-M.G., R.H., M.H.; visualization, A.-M.G.; supervision, M.H.; project administration, A.-M.G. and M.H.; funding acquisition, M.H. All authors have read and agreed to the published version of the manuscript.

References

[1]	Center for Disease Control and Prevention, Clinical alert to US healthcare facilities: global emergence of invasive infections caused by the multidrug-resistant yeast Candida auris, 2016. Available from: https://archive.cdc.gov/www_cdc_gov/fungal/candida-auris/candida-auris-alert.html
[2]	Parums DV (2022) Editorial: The World Health Organization (WHO) fungal priority pathogens list in response to emerging fungal pathogens during the COVID-19 pandemic. Med Sci Monit 28: e939088. https://doi.org/10.12659/MSM.939088
[3]	Lockhart SR (2019) Candida auris and multidrug resistance: defining the new normal. Fungal Genet Biol 131: 103243. https://doi.org/10.1016/j.fgb.2019.103243
[4]	Ademe M, Girma F (2020) Candida auris: from multidrug resistance to pan-resistant strains. Infect Drug Resist 13: 1287-1294. https://doi.org/10.2147/IDR.S249864
[5]	Larkin E, Hager C, Chandra J, et al. (2017) The emerging pathogen Candida auris: growth phenotype, virulence factors, activity of antifungals, and effect of SCY-078, a novel glucan synthesis inhibitor, on growth morphology and biofilm formation. Antimicrob Agents Ch 61: 13. https://doi.org/10.1128/aac.02396-16
[6]	Wang X, Bing J, Zheng Q, et al. (2018) The first isolate of Candida auris in China: clinical and biological aspects. Emerg Microbes Infec 7: 93. https://doi.org/10.1038/s41426-018-0095-0
[7]	Fan S, Yue H, Zheng Q, et al. (2021) Filamentous growth is a general feature of Candida auris clinical isolates. Med Mycol 59: 734-740. https://doi.org/10.1093/mmy/myaa116
[8]	Tharp B, Zheng R, Bryak G, et al. (2023) Role of microbiota in the skin colonization of Candida auris. mSphere 8: e0062322. https://doi.org/10.1128/msphere.00623-22
[9]	Grice EA, Segre JA (2011) The skin microbiome. Nat Rev Microbiol 9: 244-253. https://doi.org/10.1038/nrmicro2537
[10]	Robinson S, Robinson AH (1954) Chemical composition of sweat. Physiol Rev 34: 202-220. https://doi.org/10.1152/physrev.1954.34.2.202
[11]	Baker LB (2019) Physiology of sweat gland function: the roles of sweating and sweat composition in human health. Temperature (Austin) 6: 211-259. https://doi.org/10.1080/23328940.2019.1632145
[12]	Eix EF, Nett JE (2022) Modeling Candida auris skin colonization: mice, swine, and humans. Plos Pathog 18: e1010730. https://doi.org/10.1371/journal.ppat.1010730
[13]	Satoh K, Makimura K, Hasumi Y, et al. (2009) Candida auris sp. nov., a novel ascomycetous yeast isolated from the external ear canal of an inpatient in a Japanese hospital. Microbiol Immunol 53: 41-44. https://doi.org/10.1111/j.1348-0421.2008.00083.x
[14]	Sakamoto FH, Lopes JD, Anderson RR (2010) Photodynamic therapy for acne vulgaris: a critical review from basics to clinical practice: part I. Acne vulgaris: when and why consider photodynamic therapy?. J Am Acad Dermatol 63: 183-193. https://doi.org/10.1016/j.jaad.2009.09.056
[15]	Yang F, Xu M, Chen X, et al. (2023) Spotlight on porphyrins: classifications, mechanisms and medical applications. Biomed Pharmacother 164: 114933. https://doi.org/10.1016/j.biopha.2023.114933
[16]	Barron GA, Moseley H, Woods JA (2013) Differential sensitivity in cell lines to photodynamic therapy in combination with ABCG2 inhibition. J Photochem Photobiol B 126: 87-96. https://doi.org/10.1016/j.jphotobiol.2013.07.003
[17]	Harris DM, Werkhaven J (1987) Endogenous porphyrin fluorescence in tumors. Lasers Surg Med 7: 467-472. https://doi.org/10.1002/lsm.1900070605
[18]	Plavskii VY, Mikulich AV, Tretyakova AI, et al. (2018) Porphyrins and flavins as endogenous acceptors of optical radiation of blue spectral region determining photoinactivation of microbial cells. J Photochem Photobiol B 183: 172-183. https://doi.org/10.1016/j.jphotobiol.2018.04.021
[19]	Hessling M, Wenzel U, Meurle T, et al. (2020) Photoinactivation results of Enterococcus moraviensis with blue and violet light suggest the involvement of an unconsidered photosensitizer. Biochem Biophy Rese Co 533: 813-817. https://doi.org/10.1016/j.bbrc.2020.09.091
[20]	Gierke AM, Hessling M (2024) Photoinactivation by UVA radiation and visible light of Candida auris compared to other fungi. Photochem Photobiol Sci 23: 681-692. https://doi.org/10.1007/s43630-024-00543-4
[21]	Pallotta ML (2011) Evidence for the presence of a FAD pyrophosphatase and a FMN phosphohydrolase in yeast mitochondria: a possible role in flavin homeostasis. Yeast 28: 693-705. https://doi.org/10.1002/yea.1897
[22]	Viñas P, Balsalobre N, López-Erroz C, et al. (2004) Liquid chromatographic analysis of riboflavin vitamers in foods using fluorescence detection. J Agric Food Chem 52: 1789-1794. https://doi.org/10.1021/jf030756s
[23]	Zhao C, Liu L, Ge J, et al. (2017) Ultrasensitive determination for flavin coenzyme by using a ZnO nanorod photoelectrode in a four-electrode system. Microchim Acta 184: 2333-2339. https://doi.org/10.1007/s00604-017-2230-3
[24]	Hu X, Huang YY, Wang Y, et al. (2018) Antimicrobial photodynamic therapy to control clinically relevant biofilm infections. Front Microbiol 9: 1299. https://doi.org/10.3389/fmicb.2018.01299
[25]	Plaetzer K, Krammer B, Berlanda J, et al. (2009) Photophysics and photochemistry of photodynamic therapy: fundamental aspects. Lasers Med Sci 24: 259-268. https://doi.org/10.1007/s10103-008-0539-1
[26]	Di Meo S, Reed TT, Venditti P, et al. (2016) R Role of ROS and RNS sources in physiological and pathological conditions. Oxid Med Cell Longev 2016: 1245049. https://doi.org/10.1155/2016/1245049
[27]	Mallidi S, Anbil S, Lee S, et al. (2014) Photosensitizer fluorescence and singlet oxygen luminescence as dosimetric predictors of topical 5-aminolevulinic acid photodynamic therapy induced clinical erythema. J Biomed Opt 19: 028001. https://doi.org/10.1117/1.JBO.19.2.028001
[28]	Brasch J, Kay C (2006) Effects of repeated low-dose UVB irradiation on the hyphal growth of Candida albicans. Mycoses 49: 1-5. https://doi.org/10.1111/j.1439-0507.2005.01169.x
[29]	Cieplik F, Späth A, Leibl C, et al. (2014) Blue light kills Aggregatibacter actinomycetemcomitans due to its endogenous photosensitizers. Clin Oral Investi 18: 1763-1769. https://doi.org/10.1007/s00784-013-1151-8
[30]	Lei J, Huang J, Xin C, et al. (2023) Riboflavin targets the cellular metabolic and ribosomal pathways of Candida albicans in vitro and exhibits efficacy against oropharyngeal candidiasis. Microbiol Spectr 11: e0380122. https://doi.org/10.1128/spectrum.03801-22
[31]	Oriel S, Nitzan Y (2010) Photoinactivation of Candida albicans by its own endogenous porphyrins. Curr Microbiol 60: 117-123. https://doi.org/10.1007/s00284-009-9514-8
[32]	Meir Z, Osherov N (2018) Vitamin biosynthesis as an antifungal target. J Fungi (Basel) 4: 72. https://doi.org/10.3390/jof4020072
[33]	Koenig K, Schneckenburger H (1994) Laser-induced autofluorescence for medical diagnosis. J Fluoresc 4: 17-40. https://doi.org/10.1007/BF01876650
[34]	Yang H, Xiao X, Zhao X S, et al. (2016) Study on fluorescence spectra of thiamine and riboflavin. MATEC Web Conf 63: 03013. https://doi.org/10.1051/matecconf/20166303013
[35]	Kowalska E, Kozik A (2008) The genes and enzymes involved in the biosynthesis of thiamin and thiamin diphosphate in yeasts. Cell Mol Biol Lett 13: 271-282. https://doi.org/10.2478/s11658-007-0055-5
[36]	Maitra D, Pinsky BM, Soherawardy A, et al. (2021) Protein-aggregating ability of different protoporphyrin-IX nanostructures is dependent on their oxidation and protein-binding capacity. J Biol Chem 297: 100778. https://doi.org/10.1016/j.jbc.2021.100778
[37]	Fyrestam J, Bjurshammar N, Paulsson E, et al. (2017) Influence of culture conditions on porphyrin production in Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Photodiagn Photodyn 17: 115-123. https://doi.org/10.1016/j.pdpdt.2016.11.001
[38]	Pretlow TP, Sherman F (1967) Porphyrins and zinc porphyrins in normal and mutant strains of yeast. BBA–Gen Subjects 148: 629-644. https://doi.org/10.1016/0304-4165(67)90036-0
[39]	Phillips JD (2019) Heme biosynthesis and the porphyrias. Mol Genet Metab 128: 164-177. https://doi.org/10.1016/j.ymgme.2019.04.008
[40]	Andrawes N, Weissman Z, Pinsky M, et al. (2022) Regulation of heme utilization and homeostasis in Candida albicans. Plos Genet 18: e1010390. https://doi.org/10.1371/journal.pgen.1010390
[41]	Kornitzer D, Roy U (2020) Pathways of heme utilization in fungi. Biochim Biophys Acta Mol Cell Res 1867: 118817. https://doi.org/10.1371/journal.pgen.1010390
[42]	Harris DM (2023) Spectra of pathogens predict lethality of blue light photo-inactivation. Laser Ther J 30: 314. http://dx.doi.org/10.4081/ltj.2023.314
[43]	Bohm GC, Gandara L, Di Venosa G, et al. (2020) Photodynamic inactivation mediated by 5-aminolevulinic acid of bacteria in planktonic and biofilm forms. Biochem Pharmacol 177: 114016. https://doi.org/10.1016/j.bcp.2020.114016
[44]	Melø TB, Johnsson M (1982) In vivo porphyrin fluorescence for Propionibacterium acnes. A characterization of the fluorescing pigments. Dermatologica 164: 167-174. https://doi.org/10.1159/000250086
[45]	Gupta S, MacLean M, Anderson JG, et al. (2015) Inactivation of micro-organisms isolated from infected lower limb arthroplasties using high-intensity narrow-spectrum (HINS) light. Bone Joint J 97: 283-288. https://doi.org/10.1302/0301-620X.97B2.35154
[46]	Rosa LP, Da Silva FC, Viana MS, et al. (2016) In vitro effectiveness of 455-nm blue LED to reduce the load of Staphylococcus aureus and Candida albicans biofilms in compact bone tissue. Lasers Med Sci 31: 27-32. https://doi.org/10.1007/s10103-015-1826-2
[47]	Tsutsumi-Arai C, Arai Y, Terada-Ito C, et al. (2022) Microbicidal effect of 405-nm blue LED light on Candida albicans and Streptococcus mutans dual-species biofilms on denture base resin. Lasers Med Sci 37: 857-866. https://doi.org/10.1007/s10103-021-03323-z
[48]	Wang T, Dong J, Yin H, et al. (2020) Blue light therapy to treat candida vaginitis with comparisons of three wavelengths: an in vitro study. Lasers Med Sci 35: 1329-1339. https://doi.org/10.1007/s10103-019-02928-9
[49]	Pérez-Laguna V, García-Luque I, Ballesta S, et al. (2021) Photodynamic therapy combined with antibiotics or antifungals against microorganisms that cause skin and soft tissue infections: A planktonic and biofilm approach to overcome resistances. Pharmaceuticals 14: 603. https://doi.org/10.3390/ph14070603
[50]	Pérez-Laguna V, Gilaberte Y, Millán-Lou MI, et al. (2019) A combination of photodynamic therapy and antimicrobial compounds to treat skin and mucosal infections: a systematic review. Photochem Photobiol Sci 18: 1020-1029. https://doi.org/10.1039/c8pp00534f
[51]	Wozniak A, Rapacka-Zdonczyk A, Mutters NT, et al. (2019) Antimicrobials are a photodynamic iinactivation adjuvant for the eradication of extensively drug-resistant Acinetobacter baumannii. Front Microbiol 10: 229. https://doi.org/10.3389/fmicb.2019.00229
[52]	Ilizirov Y, Formanovsky A, Mikhura I, et al. (2018) Effect of photodynamic antibacterial chemotherapy combined with antibiotics on gram-positive and gram-negative bacteria. Molecules 23: 3152. https://doi.org/10.3390/molecules23123152

Reader Comments

Your name:*

Email:*
© 2025 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)