Influence of cobalt chloride and ferric citrate on purple non-sulfur bacteria Rhodopseudomonas yavorovii

  • S. O. Hnatush Ivan Franko National University of Lviv
  • O. D. Maslovska Ivan Franko National University of Lviv
  • S. Y. Komplikevych Ivan Franko National University of Lviv
  • I. V. Kovbasa Ivan Franko National University of Lviv
Keywords: free radical damage; lipid peroxidation; heavy metals; oxidative modification of proteins; antioxidant defense system; cobalt ions; ferric ions.


Heavy metals that enter the environment due to natural processes or industrial activities, when accumulated, have a negative impact on organisms, including microorganisms. Microorganisms have developed various adaptations to heavy metal compounds. The aim of our work was to investigate the influence of ferric citrate and cobalt (II) chloride on biomass accumulation, indicators of free radical damage and activity of enzymes of the antioxidant defense system of bacteria Rhodopseudomonas yavorovii IMV B-7620, that were isolated from the water of Yavorivske Lake (Ukraine, Lviv region), which was formed as a result of flooding of a sulfur quarry. We used cultural, photometric methods, and statistical processing of the results was performed using two-way ANOVA and factor analysis. It was found that ferric citrate at a concentration of 1–12 mM causes inhibition of the accumulation of biomass of bacteria Rh. yavorovii IMV B-7620 up to 44.7%, and cobalt (II) chloride at a concentration of 1–15 mM – up to 70.4%, compared with the control. The studied concentrations of ferric citrate and cobalt (II) chloride cause free radical damage to lipids and proteins of Rh. yavorovii IMV B-7620. As a result of two-way ANOVA we found that under the influence of ferric citrate statistically significant changes in biomass accumulation, lipid hydroperoxides and thiobarbiturate reactive species content, superoxide dismutase activity were predetermined by increasing the concentration of metal salts as well as increasing the duration of cultivation of bacteria, while the content of diene conjugates and catalase activity changed with increasing duration of cultivation. Under the influence of cobalt (II) chloride, statistically significant changes in all studied indicators were found both due to the increase in the concentration of metal salts and with increasing duration of bacterial cultivation. The studied parameters of Rh. yavorovii IMV B-7620 cells under the influence of ferric citrate and cobalt (II) chloride are combined into two factors, that explain 95.4% and 99.2% of the total data variance, respectively. Under the influence of ferric citrate, the first latent factor included diene conjugates, thiobarbiturate reactive species, carbonyl groups in proteins, which are closely linked by a direct bond and inversely related to the content of lipid hydroperoxides and catalase activity. The second latent factor included duration of cultivation of bacteria, biomass accumulation, and superoxide dismutase activity, which are inversely related to lipid hydroperoxide content and catalase activity. Under the influence of cobalt (II) chloride, the first latent factor included the content of lipid hydroperoxides, carbonyl groups in proteins, as well as catalase and superoxide dismutase activities, which are inversely related to bacterial biomass.


Asif, A., Mohsin, H., & Rehman, Y. (2021). Purple nonsulfur bacteria: An important versatile tool in biotechnology. In: De Mandal, S., & Passari, A. K. (Eds.). Recent advancement in microbial biotechnology. Academic Press. Pp. 309–337.

Ayangbenro, A. S., & Babalola, O. O. (2017). A new strategy for heavy metal polluted environments: A review of microbial biosorbents. International Journal of Environmental Research and Public Health, 14(1), 94.

Barra Caracciolo, A., & Terenzi, V. (2021). Rhizosphere microbial communities and heavy metals. Microorganisms, 9(7), 1462.

Barras, F., & Fontecave, M. (2011). Cobalt stress in Escherichia coli and Salmonella enterica: Molecular bases for toxicity and resistance. Metallomics, 3(11), 1130–1134.

Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.

Bunraksa, T., Kantachote, D., & Chaiprapat, S. (2020). The potential use of purple nonsulfur bacteria to simultaneously treat chicken slaughterhouse wastewater and obtain valuable plant growth promoting effluent and their biomass for agricultural application. Biocatalysis and Agricultural Biotechnology, 28, 101721.

Cerruti, M., Stevens, B., Ebrahimi, S., Alloul, A., Vlaeminck, S. E., & Weissbrodt, D. G. (2020). Enrichment and aggregation of purple non-sulfur bacteria in a mixed-culture sequencing-batch photobioreactor for biological nutrient removal from wastewater. Frontiers in Bioengineering and Biotechnology, 8, 1432.

Chanpiwat, P., Sthiannopkao, S., & Kim, K. W. (2010). Metal content variation in wastewater and biosludge from Bangkok’s central wastewater treatment plants. Microchemical Journal, 95(2), 326–332.

da Silva Brito, G. F., Oliveira, R., Grisolia, C. K., Guirra, L. S., Weber, I. T., & de Almeida, F. V. (2019). Evaluation of advanced oxidative processes in biodiesel wastewater treatment. Journal of Photochemistry and Photobiology A: Chemistry, 375, 85–90.

Delgado-Sarmiento, P. (2020). Bioremediation of chromium VI by applying Rhodopseudomonas palustris in industrial effluents coming from tannery. Revista Boliviana de Química, 37(1), 21–27.

Dulay, H., Tabares, M., Kashefi, K., & Reguera, G. (2020). Cobalt resistance via detoxification and mineralization in the iron-reducing bacterium Geobacter sulfurreducens. Frontiers in Microbiology, 11, 2992.

Gari, M. K., Lemke, P., Lu, K. H., Laudadio, E. D., Henke, A. H., Green, C. M., Pho, T., Hoang, K. N. L., Murphy, C. J., Hamers, R. J., & Feng, Z. V. (2021). Dynamic aqueous transformations of lithium cobalt oxide nanoparticle induce distinct oxidative stress responses of B. subtilis. Environmental Science: Nano, 8, 1614–1627.

Garimella, S., Kugle, K. R., Kssoju, A., & Merugu, R. (2017). Current status on single cell protein (SCP) production from photosynthetic purple non sulfur bacteria. Journal of Chemical and Pharmaceutical Sciences, 10, 915–922.

Grune, T., Jung, T., Merker, K., & Davies, K. J. (2004). Decreased proteolysis caused by protein aggregates, inclusion bodies, plaques, lipofuscin, ceroid, and ‘aggresomes’ during oxidative stress, aging, and disease. The International Journal of Biochemistry and Cell Biology, 36(12), 2519–2530.

Huang, H., Li, B., Li, J., Zhang, P., Yu, W., Zhao, N., Guo, G., & Young, B. (2019). Influence of process parameters on the heavy metal (Zn2+, Cu2+ and Cr3+) content of struvite obtained from synthetic swine wastewater. Environmental Pollution, 245, 658–665.

Kapahi, M., & Sachdeva, S. (2019). Bioremediation options for heavy metal pollution. Journal of Health and Pollution, 9(24), 191203.

Kaushik, A., Basu, S., Batra, V. S., & Balakrishnan, M. (2018). Fractionation of sugarcane molasses distillery wastewater and evaluation of antioxidant and antimicrobial characteristics. Industrial Crops and Products, 118, 73–80.

Kumar, V., Mishra, R. K., Kaur, G., & Dutta, D. (2017). Cobalt and nickel impair DNA metabolism by the oxidative stress independent pathway. Metallomics, 9(11), 1596–1609.

Li, C., Yu, Y., Fang, A., Feng, D., Du, M., Tang, A., Chen, S., & Li, A. (2022). Insight into biosorption of heavy metals by extracellular polymer substances and the improvement of the efficacy: A review. Letters in Applied Microbiology, 2022, in press.

Li, D., Liu, J., Wang, S., & Cheng, J. (2020). Study on coal water slurries prepared from coal chemical wastewater and their industrial application. Applied Energy, 268, 114976.

Liu, X., Liu, H., Wu, W., Zhang, X., Gu, T., Zhu, M., & Tan, W. (2020). Oxidative stress induced by metal ions in bioleaching of LiCoO2 by an acidophilic microbial consortium. Frontiers in Microbiology, 10, 3058.

Lu, H., Zhang, G., He, S., Zhao, R., & Zhu, D. (2021). Purple non-sulfur bacteria technology: A promising and potential approach for wastewater treatment and bioresources recovery. World Journal of Microbiology and Biotechnology, 37(9), 1–15.

Malovanyy, M., Moroz, O., Hnatush, S., Maslovska, O., Zhuk, V., Petrushka, I., Nykyforov, V., & Sereda, A. (2019). Perspective technologies of the treatment of the wastewaters with high content of organic pollutants and ammoniacal nitrogen. Journal of Ecological Engineering, 20(2), 8–15.

Mohammed, A. S., Kapri, A., & Goel, R. (2011). Heavy metal pollution: Source, impact, and remedies. In: Khan, M. S., Zaidi, A., Goel, R., & Musarrat, J. (Eds.). Biomanagement of metal-contaminated soils. Springer, Dordrecht. Pp. 1–28.

Mohsin, H., Asif, A., & Rehman, Y. (2019). Anoxic growth optimization for metal respiration and photobiological hydrogen production by arsenic‐resistant Rhodopseudomonas and Rhodobacter species. Journal of Basic Microbiology, 59(12), 1208–1216.

Monroy, I., & Buitrón, G. (2020). Production of polyhydroxybutyrate by pure and mixed cultures of purple non-sulfur bacteria: A review. Journal of Biotechnology, 317, 39–47.

Montiel-Corona, V., & Buitrón, G. (2021). Polyhydroxyalkanoates from organic waste streams using purple non-sulfur bacteria. Bioresource Technology, 323, 124610.

Prabhakaran, P., Ashraf, M. A., & Aqma, W. S. (2016). Microbial stress response to heavy metals in the environment. RSC Advances, 6(111), 109862–109877.

Presentato, A., Piacenza, E., Cappelletti, M., & Turner, R. J. (2019). Interaction of Rhodococcus with metals and biotechnological applications. In: Alvarez, H. M. (Eds.). Biology of Rhodococcus. Springer, Cham. Pp. 333–357.

Rajyalaxmi, K., Merugu, R., Girisham, S., & Reddy, S. M. (2019). Chromate reduction by purple non sulphur phototrophic bacterium Rhodobacter sp. GSKRLMBKU–03 isolated from pond water. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences, 89(1), 259–265.

Reaksputi, R., Boonprab, K., Tunkijjanukij, S., & Salaenoi, J. (2019). Carotenoid production at various salinities in bacterium Rhodopseudomonas palustris. Agriculture and Natural Resources, 53(5), 500–505.

Sagir, E., & Alipour, S. (2021). Photofermentative hydrogen production by immobilized photosynthetic bacteria: Current perspectives and challenges. Renewable and Sustainable Energy Reviews, 141, 110796.

Simonsen, L. O., Harbak, H., & Bennekou, P. (2012). Cobalt metabolism and toxicology – a brief update. Science of the Total Environment, 432, 210–215.

Singh, A., Parihar, P., Singh, R., & Prasad, S. M. (2016). An assessment to show toxic nature of beneficial trace metals: Too much of good thing can be bad. International Journal of Current Multidisciplinary Studies, 2, 141–144.

Tarabas, O. V., Hnatush, S. O., Tashyrev, O. B., Hovorukha, V. M., Havryliuk, O. A., Moroz, O. M., & Halushka, A. A. (2021). Production of hydrogen by purple non-sulfur bacteria Rhodopseudomonas yavorovii IMV B-7620. Mіkrobіologіchnij Zhurnal, 83(5), 19–29.

Vasylіv, O. M., & Hnatush, S. O. (2013). Vplyv spoluk perehіdnyh metalіv na aktivnіst’ superoksyddysmutazy sіrkovіdnovliuvanyh bakterіj Desulfuromonas acetoxidans [Influence of transition metal compounds on superoxide dismutase activity of sulfur reducing Desulfuromonas acetoxidans bacteria]. Mіkrobіologіchnij Zhurnal, 75(2), 37–44 (in Ukrainian).

Wang, G., Conover, R. C., Benoit, S., Olczak, A. A., Olson, J. W., Johnson, M. K., & Maier, R. J. (2004). Role of a bacterial organic hydroperoxide detoxification system in preventing catalase inactivation. Journal of Biological Chemistry, 279(50), 51908–51914.

Wang, H., Yang, A., Zhang, G., Ma, B., Meng, F., Peng, M., & Wang, H. (2017). Enhancement of carotenoid and bacteriochlorophyll by high salinity stress in photosynthetic bacteria. International Biodeterioration and Biodegradation, 121, 91–96.

Yang, Y., Hu, Y., Duan, A., Wang, X. C., Ngo, H. H., & Li, Y. Y. (2021). Characterization of preconcentrated domestic wastewater toward efficient bioenergy recovery: Applying size fractionation, chemical composition and biomethane potential assay. Bioresource Technology, 319, 124144.

Yin, K., Wang, Q., Lv, M., & Chen, L. (2019). Microorganism remediation strategies towards heavy metals. Chemical Engineering Journal, 360, 1553–1563.