INTRODUCTION: Staphylococcus capitis naturally colonizes the human skin but as an opportunistic pathogen, it can also cause biofilm-associated infections and bloodstream infections in newborns. Previously, we found that two strains from the subspecies S. capitis subsp. capitis produce yellow carotenoids despite the initial species description, reporting this subspecies as non-pigmented. In Staphylococcus aureus, the golden pigment staphyloxanthin is an important virulence factor, protecting cells against reactive oxygen species and modulating membrane fluidity. METHODS: In this study, we used two pigmented (DSM 111179 and DSM 113836) and two non-pigmented S. capitis subsp. capitis strains (DSM 20326T and DSM 31028) to identify the pigment, determine conditions under which pigment-production occurs and investigate whether pigmented strains show increased resistance to ROS and temperature stress. RESULTS: We found that the non-pigmented strains remained colorless regardless of the type of medium, whereas intensity of pigmentation in the two pigmented strains increased under low nutrient conditions and with longer incubation times. We were able to detect and identify staphyloxanthin and its derivates in the two pigmented strains but found that methanol cell extracts from all four strains showed ROS scavenging activity regardless of staphyloxanthin production. Increased survival to cold temperatures (-20°C) was detected in the two pigmented strains only after long-term storage compared to the non-pigmented strains. CONCLUSION: The identification of staphyloxanthin in S. capitis is of clinical relevance and could be used, in the same way as in S. aureus, as a possible target for anti-virulence drug design.

Department of Algorithmic Bioinformatics Justus Liebig University Giessen Giessen Germany

Department of Bioinformatics and Systems Biology Justus Liebig University Giessen Giessen Germany

Department of Microbiology Faculty of Medicine Masaryk University and St Anne's University Hospital Brno Czechia

Department of Radiation Biology Institute of Aerospace Medicine German Aerospace Center Cologne Germany

Institute of Nutritional and Food Sciences Food Sciences University of Bonn Bonn Germany

Institute of Scientific Instruments of the Czech Academy of Sciences Brno Czechia

Zobrazit více v PubMed

Asker D., Beppu T., Ueda K. (2007). Unique diversity of carotenoid-producing bacteria isolated from Misasa, a radioactive site in Japan. Appl. Microbiol. Biotechnol. 77, 383–392. doi: 10.1007/s00253-007-1157-8, PMID: PubMed DOI

Ayala O. D., Wakeman C. A., Pence I. J., Gaddy J. A., Slaughter J. C., Skaar E. P., et al. (2018). Drug-resistant PubMed DOI PMC

Bannerman T. L., Kloos W. E. (1991). PubMed DOI

Beard-Pegler M. A., Stubbs E., Vickery A. M. (1988). Observations on the resistance to drying of staphylococcal strains. J. Med. Microbiol. 26, 251–255. doi: 10.1099/00222615-26-4-251, PMID: PubMed DOI

Bernatova S., Samek O., Pilat Z., Sery M., Jezek J., Jakl P., et al. (2013). Following the mechanisms of bacteriostatic versus bactericidal action using Raman spectroscopy. Molecules 18, 13188–13199. doi: 10.3390/molecules181113188, PMID: PubMed DOI PMC

Breithaupt D. E., Schwack W., Wolf G., Hammes W. P. (2001). Characterization of the triterpenoid 4,4′-diaponeurosporene and its isomers in food-associated bacteria. Eur. Food Res. Technol. 213, 231–233. doi: 10.1007/s002170100358 DOI

Bull A. T. (1970). Inhibition of polysaccharases by melanin: enzyme inhibition in relation to mycolysis. Arch. Biochem. Biophys. 137, 345–356. doi: 10.1016/0003-9861(70)90448-0, PMID: PubMed DOI

Cameron D. R., Jiang J. H., Hassan K. A., Elbourne L. D., Tuck K. L., Paulsen I. T., et al. (2015). Insights on virulence from the complete genome of PubMed DOI PMC

Chong C. E., Bengtsson R. J., Horsburgh M. J. (2022). Comparative genomics of PubMed DOI PMC

Clauditz A., Resch A., Wieland K. P., Peschel A., Götz F. (2006). Staphyloxanthin plays a role in the fitness of PubMed DOI PMC

Cui B., Smooker P. M., Rouch D. A., Daley A. J., Deighton M. A. (2013). Differences between two clinical PubMed DOI PMC

De Maesschalck R., Jouan-Rimbaud D., Massart D. L. (2000). The Mahalanobis distance. Chemom. Intell. Lab. Syst. 50, 1–18. doi: 10.1016/S0169-7439(99)00047-7, PMID: DOI

Decalonne M., Dos Santos S., Gimenes R., Goube F., Abadie G., Aberrane S., et al. (2020). PubMed DOI PMC

Dieckmann M. A. B. S., Nkouamedjo-Fankep R. C., Hanel P. H. G., Jelonek L., Blom J., Goesmann A. (2021). EDGAR3.0: comparative genomics and phylogenomics on a scalable infrastructure. Nucleic Acids Res. 49, W185–W192. doi: 10.1093/nar/gkab341, PMID: PubMed DOI PMC

Dieser M., Greenwood M., Foreman C. M. (2010). Carotenoid pigmentation in Antarctic heterotrophic bacteria as a strategy to withstand environmental stresses. Arct. Antarct. Alp. Res. 42, 396–405. doi: 10.1657/1938-4246-42.4.396 DOI

Edgar R. C. (2004). MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 32, 1792–1797. doi: 10.1093/nar/gkh340, PMID: PubMed DOI PMC

El-Agamey A., Lowe G. M., McGarvey D. J., Mortensen A., Phillip D. M., Truscott T. G., et al. (2004). Carotenoid radical chemistry and antioxidant/pro-oxidant properties. Arch. Biochem. Biophys. 430, 37–48. doi: 10.1016/j.abb.2004.03.007, PMID: PubMed DOI

Etzbach L., Pfeiffer A., Weber F., Schieber A. (2018). Characterization of carotenoid profiles in goldenberry ( PubMed DOI

Fong N., Burgess M., Barrow K., Glenn D. (2001). Carotenoid accumulation in the psychrotrophic bacterium PubMed DOI

Gilchrist C. L. M., Chooi Y.-H. (2021). clinker & clustermap.js: automatic generation of gene cluster comparison figures. Bioinformatics 37, 2473–2475. doi: 10.1093/bioinformatics/btab007, PMID: PubMed DOI

Kaiser P., Surmann P., Vallentin G., Fuhrmann H. (2007). A small-scale method for quantitation of carotenoids in bacteria and yeasts. J. Microbiol. Methods 70, 142–149. doi: 10.1016/j.mimet.2007.04.004, PMID: PubMed DOI

Kim M., Jung D. H., Hwang C. Y., Siziya I. N., Park Y. S., Seo M. J. (2023). 4,4′-diaponeurosporene production as C30 carotenoid with antioxidant activity in recombinant PubMed DOI

Kim S. H., Kim M. S., Lee B. Y., Lee P. C. (2016). Generation of structurally novel short carotenoids and study of their biological activity. Sci. Rep. 6, 1–11. doi: 10.1038/srep21987, PMID: PubMed DOI PMC

Linden M., Flegler A., Feuereisen M. M., Weber F., Lipski A., Schieber A. (2023). Effects of flavonoids on membrane adaptation of food-associated bacteria. Biochim. Biophys. Acta 1865:184137. doi: 10.1016/j.bbamem.2023.184137, PMID: PubMed DOI

Liu G. Y., Essex A., Buchanan J. T., Datta V., Hoffman H. M., Bastian J. F., et al. (2005). PubMed DOI PMC

López G. D., Suesca E., Álvarez-Rivera G., Rosato A. E., Ibáñez E., Cifuentes A., et al. (2021). Carotenogenesis of PubMed DOI

Madeira F., Pearce M., Tivey A. R. N., Basutkar P., Lee J., Edbali O., et al. (2022). Search and sequence analysis tools services from EMBL-EBI in 2022. Nucleic Acids Res. 50, W276–W279. doi: 10.1093/nar/gkac240, PMID: PubMed DOI PMC

Marshall J. H., Wilmoth G. J. (1981). Pigments of PubMed DOI PMC

Merrikh C. N., Merrikh H. (2018). Gene inversion potentiates bacterial evolvability and virulence. Nat. Commun. 9:4662. doi: 10.1038/s41467-018-07110-3 PubMed DOI PMC

Mishra N. N., Liu G. Y., Yeaman M. R., Nast C. C., Proctor R. A., McKinnell J., et al. (2011). Carotenoid-related alteration of cell membrane fluidity impacts PubMed DOI PMC

Mlynarikova K., Samek O., Bernatova S., Ruzicka F., Jezek J., Haronikova A., et al. (2015). Influence of culture media on microbial fingerprints using Raman spectroscopy. Sensors 15, 29635–29647. doi: 10.3390/s151129635, PMID: PubMed DOI PMC

Moore J., Yu L. (2007). “Methods for antioxidant capacity estimation of wheat and wheat-based food products” in Wheat antioxidants. ed. Yu L. (Hoboken, New Jersey, United States: John Wiley & Sons, Inc.), 118–172.

Mora M., Wink L., Kögler I., Mahnert A., Rettberg P., Schwendner P., et al. (2019). Space station conditions are selective but do not alter microbial characteristics relevant to human health. Nat. Commun. 10:3990. doi: 10.1038/s41467-019-11682-z PubMed DOI PMC

Natsis N. E., Cohen P. R. (2018). Coagulase-negative PubMed DOI

Palma M., Cheung A. L. (2001). ς PubMed DOI PMC

Pannu M. K., Hudman D. A., Sargentini N. J., Singh V. K. (2019). Role of SigB and staphyloxanthin in radiation survival of PubMed DOI

Pelz A., Wieland K.-P., Putzbach K., Hentschel P., Albert K., Götz F. (2005). Structure and biosynthesis of staphyloxanthin from PubMed DOI

Rebrošová K., Šiler M., Samek O., Růžička F., Bernatová S., Ježek J., et al. (2017). Differentiation between PubMed DOI

Renishaw (2003). Technical note: Renishaw’s EasyConfocal Raman method SPD/TN/076. Wotton-under-Edge. Rehnishaw PLC

Sajjad W., Din G., Rafiq M., Iqbal A., Khan S., Zada S., et al. (2020). Pigment production by cold-adapted bacteria and fungi: colorful tale of cryosphere with wide range applications. Extremophiles 24, 447–473. doi: 10.1007/s00792-020-01180-2, PMID: PubMed DOI PMC

Salamzade R., Cheong J. Z. A., Sandstrom S., Swaney M. H., Stubbendieck R. M., Starr N. L., et al. (2023). Evolutionary investigations of the biosynthetic diversity in the skin microbiome using PubMed DOI PMC

Schwengers O., Jelonek L., Dieckmann M. A., Beyvers S., Blom J., Goesmann A. (2021). Bakta: rapid and standardized annotation of bacterial genomes via alignment-free sequence identification. Microb. Genom. 7:000685. doi: 10.1099/mgen.0.000685, PMID: PubMed DOI PMC

Seel W., Baust D., Sons D., Albers M., Etzbach L., Fuss J., et al. (2020). Carotenoids are used as regulators for membrane fluidity by PubMed DOI PMC

Sendra J. M., Sentandreu E., Navarro J. L. (2006). Reduction kinetics of the free stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH DOI

Siems K., Runzheimer K., Rehm A., Schwengers O., Heidler von Heilborn D., Kaser L., et al. (2022). Phenotypic and genomic assessment of the potential threat of human spaceflight-relevant PubMed DOI PMC

Siziya I. N., Hwang C. Y., Seo M.-J. (2022). Antioxidant potential and capacity of microorganism-sourced C30 carotenoids—a review. Antioxidants 11:1963. doi: 10.3390/antiox11101963, PMID: PubMed DOI PMC

Sobisch L.-Y., Rogowski K. M., Fuchs J., Schmieder W., Vaishampayan A., Oles P., et al. (2019). Biofilm forming antibiotic resistant gram-positive pathogens isolated from surfaces on the international Space Station. Front. Microbiol. 10:543. doi: 10.3389/fmicb.2019.00543, PMID: PubMed DOI PMC

Verma S. P., Philippot J. R., Bonnet B., Sainte-Marie J., Moschetto Y., Wallach D. F. (1984). Resonance Raman spectra of beta-carotene in native and modified low-density lipoprotein. Biochem. Biophys. Res. Commun. 122, 867–875. doi: 10.1016/S0006-291X(84)80114-X, PMID: PubMed DOI

ViroxTechnologies (2018). AHP goes beyond. Available at: https://virox.com/making-news/virox-corp-news/ahp-goes-beyond/ (Accessed August 31, 2023).

Watanabe S., Aiba Y., Tan X.-E., Li F.-Y., Boonsiri T., Thitiananpakorn K., et al. (2018). Complete genome sequencing of three human clinical isolates of PubMed DOI PMC

Wieland B., Feil C., Gloria-Maercker E., Thumm G., Lechner M., Bravo J. M., et al. (1994). Genetic and biochemical analyses of the biosynthesis of the yellow carotenoid 4,4′-diaponeurosporene of PubMed DOI PMC

Wirth T., Bergot M., Rasigade J.-P., Pichon B., Barbier M., Martins-Simoes P., et al. (2020). Niche specialization and spread of PubMed DOI

Xue L., Chen Y. Y., Yan Z., Lu W., Wan D., Zhu H. (2019). Staphyloxanthin: a potential target for antivirulence therapy. Infect. Drug Resist. 12:2151. doi: 10.2147/IDR.S193649 PubMed DOI PMC

Yao Z. Y., Qi J. H. (2016). Comparison of antioxidant activities of melanin fractions from chestnut Shell. Molecules 21:487. doi: 10.3390/molecules21040487, PMID: PubMed DOI PMC

Life in the brine of Lunenburg, Germany: unveiling microorganisms associated with Zechstein salt deposits