Evaluating the Sulfur Oxidation Capability of a Rhodopseudomonas palustris Strain by Gene and Enzyme Analyses for Potential Applications in Environmental Bioremediation
Main Article Content
Abstract
Sulfur-oxidizing bacteria are capable of oxidizing various sulfur compounds including sulfide and thiosulfate in the environment. Hydrogen sulfide (H2S), a product of anaerobic decomposition, is highly corrosive. With the typical smell of rotten eggs, the toxic gas H2S causes many harmful effects on the environment and human health. Sulfide:quinone oxidoreductase enzyme (SQR) produced by sulfur-oxidizing bacteria plays an important role in the sulfide oxidation process, contributing to minimizing sulfide toxicity in the environment. In this study, the presence of sqr gene and SQR enzyme in the photosynthetic purple bacterium Rhodopseudomonas palustris (R. palustris) isolated from domestic wastewater was determined by gene amplification (PCR) using specific primer pairs following by sequencing and denaturing SDS-PAGE method combined with sulfide oxidase assay that measures BaSO4 turbidity, respectively. The results showed that the sqr gene has 1254 bp in length, encoding for the SQR protein containing 417 amino acids with a molecular weight of 45.87 kDa and locating on the cell membrane. The enzyme operated optimally under the condition of pH 6.5 and temperature at 30 °C with sulfide oxidation activity recorded up to 20-21 U/ml. The study has initially shown the ability of the R. palustris bacterial strain to oxidize sulfide for further research using sulfur-oxidizing bacteria applied in environmental treatment.
References
Journal of Cleaner Production , Vol. 312, 2021, pp. 127746, https://doi.org/10.1016/j.jclepro.2021.127746.
[2] D. Pokorna, J. Zabranska, Sulfur-oxidizing Bacteria in Environmental Technology, Biotechnology Advances, Vol. 33, No. 6, 2015, pp. 1246-1259, https://doi.org/10.1016/j.biotechadv.2015.02.007.
[3] T. Berben, L. Overmars, D. Y. Sorokin, G. Muyzer, Diversity and Distribution of Sulfur
Oxidation-related Genes in Thioalkalivibrio, a Genus of Chemolithoautotrophic and
Haloalkaliphilic Sulfur-Oxidizing Bacteria, Frontiers in Microbiology, Vol. 10, 2019, https://doi.org/10.3389/fmicb.2019.00160.
[4] P. M. Nguyen, P. T. Do, Y. B. Pham, T. O. Doan, X. C. Nguyen et al., Roles, Mechanism of Action, and Potential Applications of Sulfur-Oxidizing Bacteria for Environmental Bioremediation, Science of the Total Environment, Vol. 852, 2022, https://doi.org/10.1016/j.scitotenv.2022.158203.
[5] C. Griesbeck, G. Hauska, M. Schütz. Biological Sulfide Oxidation: Sulfide-Quinone Reductase (SQR), the Primary Reaction, Recent Research Developments in Microbiology Vol. 4, 2000.
[6] J. A. Brito, F. L. Sousa, M. Stelter, T. M. Bandeiras, C. Vonrhein, M. Teixeira, M. Pereira, M. Archer, Structural and Functional Insights into Sulfide: Quinone Oxidoreductase, Biochemistry, Vol. 48, No. 24, 2009, pp. 5613-5622, https://doi.org/10.1021/bi9003827.
[7] Y. Shahak, C. Klughammer, U. Schreiber, E. Padan, I. Herrman, G. Hauska, Sulfide-quinone and Sulfide-Cytochrome Reduction in Rhodobacter capsulatus. Photosynthesis Research, Vol. 39, 1994, pp. 175-181, https://doi.org/10.1007/BF00029384.
[8] M. M. Cherney, Y. Zhang, M. N. G. James, J. H. Weiner, Structure-Activity Characterization of Sulfide:quinone Oxidoreductase Variants, Journal of Structural Biology, Vol. 178, No. 3, 2012,
pp. 319-328, https://doi.org/10.1016/j.jsb.2012.04.007.
[9] M. Li, P. Ning, Y. Sun, J. Luo, J. Yang, Characteristics and Application of Rhodopseudomonas palustris as a Microbial Cell Factory, Frontiers in Bioengineering and Biotechnology, Vol. 10, 2022,
https://doi. org/10.3389/fbioe.2022.897003.
[10] P. Wu, Z. Chen, Y. Zhang, Y. Wang, F. Zhu, B. Cao, Y. Wu, N. Li, Rhodopseudomonas palustris Wastewater Treatment: Cyhalofop-butyl Removal, Biochemicals Production and Mathematical Model Establishment, Bioresource Technology, Vol. 282, 2019, pp. 390-397, https://doi.org/10.1016/j.biortech.2018.11.087
[11] Z. Peirong, L. Wei, Use of Fluidized Bed Biofilter and Immobilized Rhodopseudomonas palustris for Ammonia Removal and Fish Health Maintenance in a Recirculation Aquaculture System, Aquaculture Research, Vol. 44, No. 3, 2013, pp. 327-334, https://doi.org/10.1111/j.1365-2109.2011.03038.x.
[12] D. B. Minh, P. T. Ha, Isolation and Selection of Purple Non-Sulfur Bacteria for Nutrient Rich Biomass Production from Wastes, VNU Journal of Science: Natural Sciences and Technology,
Vol. 37, No. 2, 2021, pp. 24-34, https://doi.org/10.25073/2588-1140/vnunst.5121.
[13] K. Wilson, Preparation of Genomic DNA from Bacteria, Current Protocols in Molecular Biology Vol. 56, 2001, https://doi.org/10.1002/0471142727.mb0204s56.
[14] M. G. Klotz, S. W. Hutcheson, Multiple Periplasmic Catalases in Phytopathogenic Strains of Pseudomonas syringae, Applied and Environmental Microbiology, Vol. 58, No. 8, 1992, pp. 2468-2473, https://doi.org/10.1128/aem.58.8.2468-2473.1992.
[15] T. Hirano, H. Kurosawa, K. Nakamura, Y. Amano, Simultaneous Removal of Hydrogen Sulphide and Trimethylamine by a Bacterial Deodorant, Journal of Fermentation and Bioengineering, Vol. 81,
No. 4, 1996, pp. 337-342, https://doi.org/10.1016/0922-338X(96)80587-3.
[16] A. Kolmert, P. Wikström. P. K. B. Hallberg, A fast and Simple Turbidimetric Method for the Determination of Sulfate in Sulfate-reducing Bacterial Cultures, Journal of Microbiological Methods, Vol. 41, No. 3, 2000, pp. 179-184, https://doi.org/10.1016/S0167-7012(00)00154-8.
[17] C. Griesbeck, M. Schütz, T. Schödl, S. Bathe, L. Nausch, N. Mederer, M. Vielreicher, G. Hauska, Mechanism of Sulfide-quinone Reductase Investigated Using Site-directed Mutagenesis and Sulfur Analysis, Biochemistry, Vol. 41, No. 39, 2002, pp. 11552-11565, https://doi.org/10.1021/bi026032b.
[18] C. Schimek, E. Egger, C. Tauer, G. Striedner,
C. Brocard, M. C. Puschmann, R. Hahn, Extraction of Recombinant Periplasmic Proteins under Industrially Relevant Process Conditions: Selectivity and Yield Strongly Depend on Protein Titer and Methodology. Biotechnology Progress, Vol. 36, No. 5, 2020, pp. e2999, https://doi.org/10.1002/btpr.2999.
[19] B. Arieli, Y. Shahak, D. Taglicht, G. Hauska, E. Padan, Purification and Characterization of Sulfide-quinone Reductase, a Novel Enzyme Driving Anoxygenic Photosynthesis in Oscillatoria limnetica, Journal of Biological Chemistry, Vol. 269, No. 8, 1994, pp. 5705-5711, https://doi.org/10.1016/S0021-9258(17)37518-X.