Abstract
Microorganisms isolated from contaminated areas play an important role in bioremediation processes. They promote heavy metal removal from the environment by adsorbing ions onto the cell wall surface, accumulating them inside the cells, or reducing, complexing, or precipitating these substances in the environment. Microorganism-based bioremediation processes can be highly efficient, low-cost and have low environmental impact. Thus, the present study aimed to select Pb2+-resistant bacteria and evaluate the growth rate, biological activity, and the presence of genes associated with metal resistance. Serratia marcescens CCMA 1010, that was previously isolated from coffee processing wastewater, was selected since was able to growth in Pb2+ concentrations of up to 4.0 mM. The growth rate and generation time did not differ from those of the control (without Pb2+), although biological activity decreased in the first hour of exposure to these ions and stabilized after this period. The presence of the zntR, zntA and pbrA genes was analysed, and only zntR was detected. The zntR gene encodes a protein responsible for regulating the production of ZntA, a transmembrane protein that facilitates Pb2+ extrusion out of the cell. S. marcescens CCMA 1010 demonstrated a potential for use as bioindicator that has potential to be used in bioremediation processes due to its resistance to high concentrations of Pb2+, ability to grow until 24 h of exposure, and possession of a gene that indicates the existence of mechanisms associated with resistance to lead (Pb2+).
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Not applicable.
Code availability
Not applicable.
References
Ahemad M, Malik A (2011) Bioaccumulation of heavy metals by zinc resistant bacteria isolated from agricultural soils irrigated with wastewater. Bacteriol J 2:12–21. https://doi.org/10.3923/bj.2012.12.21
Bakirdere S, Bolucek C, Yaman M (2016) Determination of contamination levels of pb, cd, cu, ni and mn caused by former lead mining gallery. Environ Monit Assess 188(132):1–7. https://doi.org/10.1007/s10661-016-5134-5
Bakshi S, Leoncini E, Baker C, Cañas-Duarte SJ, Okumus B, Paulsson J (2021) Tracking bacterial lineages in complex and dynamic environments with applications for growth control and persistence. Nat Microbiol 6:783–791. https://doi.org/10.1038/s41564-021-00900-4
Becerra-Castro C, Machado RA, Vaz-Moreira I, Manaia CM (2015) Assessment of copper and zinc salts as selectors of antibiotic resistance in gram-negative bacteria. Sci Total Enviro 530–531:367–372. https://doi.org/10.1016/j.scitotenv.2015.05.102
Benmessaoud IR, Mahul-Mellier AL, Horváth E, Maco B, Spina M, Lashuel HA, Forro L (2016) Health hazards of methylammonium lead iodide based perovskites: cytotoxicity studies. Toxicol Res 5:407–419. https://doi.org/10.1039/c5tx00303b
Binet MRB, Poole RK (2000) Cd(II), Pb(II) and Zn(II) ions regulate expression of the metal-transporting P-type ATPase ZntA in Escherichia coli. FEBS Lett 473(1):67–70. https://doi.org/10.1016/S0014-5793(00)01509-X
Brocklehurst KR, Hobman JL, Blank L, Marshal SJ, Brown NL, Morby AP (1999) ZntR is a Zn ( II ) -responsive MerR-like transcriptional regulator of zntA in Escherichia coli. Mol Microbiol 31:893–902
Cai X, Zheng X, Zhang D, Iqbal W, Liu C, Yang B, Zhao X, Lu X, Mao Y (2019) Microbial characterization of heavy metal resistant bacterial strains isolated from an electroplating wastewater treatment plant. Ecotoxicol Environ Saf 181:472–480. https://doi.org/10.1016/j.ecoenv.2019.06.036
Chandrangsu P, Rensing C, Helmann JD (2017) Metal homeostasis and resistance in bacteria. Nat Rev Microbiol 15(6):338–350. https://doi.org/10.1038/nrmicro.2017.15
Chen WM, Wu CH, James EK, Chang JS (2008) Metal biosorption capability of Cupriavidus taiwanensis and its effects on heavy metal removal by nodulated Mimosa pudica. J Harzad Mater 151:364–371. https://doi.org/10.1016/j.jhazmat.2007.05.082
Chen Z, Pan X, Chen H, Guan X, Lin Z (2016) Biomineralization of Pb (II) into Pb-hydroxyapatite induced by Bacillus cereus 12–2 isolated from Lead–Zinc mine tailings. J Harzad Mater 301:531–537. https://doi.org/10.1016/j.jhazmat.2015.09.023
Chen B, Fang L, Yan X, Zhang A, Chen P, Luan T, Hu L, Jiang G (2019) A unique Pb-binding flagellin as an effective remediation tool for Pb contamination in aquatic environment. J Hazard Mater 363(5):34–40. https://doi.org/10.1016/j.jhazmat.2018.10.004
Cristani M, Naccari C, Nostro A, Pizzimenti A, Trombetta D, Pizzimenti F (2012) Possible use of Serratia marcescens in toxic metal biosorption (removal). Environ Sci Pollut Res 19(1):161–168. https://doi.org/10.1007/s11356-011-0539-8
Drewniak L, Ciezkowska M, Radlinska M, Sklodowska A (2015) Construction of the recombinant broad-host-range plasmids providing their bacterial hosts arsenic resistance and arsenite oxidation ability. J Biotechnol 196–197:42–51. https://doi.org/10.1016/j.jbiotec.2015.01.013
El Aafi N, Brhada F, Dary M, Maltouf F, Pajuelo E (2012) Rhizostabilization of metals in soils using Lupinus luteus inoculated with the metal resistant rhizobacterium Serratia SP. MSMC541. Int J Phytoremediat 14(3):261–274. https://doi.org/10.1080/15226514.2011.604693
Elizabeth CN, Victoria MY, Nkechi EE, Godwin BO (2017) Isolation and characterization of heavy metal tolerant bacteria from Panteka stream, Kaduna, Nigeria and their potential for bioremediation. Afr J Biotechnol 16(1):32–40. https://doi.org/10.5897/AJB2016.15676
Ferreira DF (2014) Sisvar: a Guide for its Bootstrap procedures in multiple comparisons. Ciênc Agrotec 38(2):109–112. https://doi.org/10.1590/S1413-70542014000200001
Gefen O, Fridman O, Ronin I, Balaban NQ (2014) Direct observation of single stationary-phase bacteria reveals a surprisingly long period of constant protein production activity. Proc Natl Acad Sci USA 111(1):556–561. https://doi.org/10.1073/pnas.1314114111
Guo H, Luo S, Chen L, Xia X, Xi Q, Wei W, Zeng G, Liu C, Wan Y, Chen J, He Y (2010) Bioremediation of heavy metals by growing hyperaccumulaor endophytic bacterium Bacillus sp. L14. Bioresour Technol 101:8599–8605. https://doi.org/10.1016/j.biortech.2010.06.085
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symposium Series, vol 41, pp 95–98. http://www.mbio.ncsu.edu/BioEdit/bioedit.html
Helmann JD, Soonsanga S, Gabriel S (2007) Metalloregulators: arbiters of metal sufficiency. In: Nies DH, Silver S (eds) Molecular microbiology of heavy metals, vol 6, 1st edn. Springer-, Berlin, pp 37–71. https://www.springer.com/gp/book/9783540697701#
Hynninen A, Touzé T, Pitkänen L, Mengin-Lecreulx D, Virta M (2009) An efflux transporter PbrA and a phosphatase PbrB cooperate in a lead-resistance mechanism in bacteria. Mol Microbiol 74(2):384–394. https://doi.org/10.1111/j.1365-2958.2009.06868.x
Huang F, Guo CL, Li GN, Yi XY, Zhu LD, Dang Z (2014) Bioaccumulation characterization of cadmium by growing Bacillus cereus RC-1 and its mechanism. Chemosphere 109:134–142. https://doi.org/10.1016/j.chemosphere.2014.01.066
Iguchi A, Nagaya Y, Pradel E, Ooka T, Ogura Y, Katsura K, Kurokawa K, Oshima K, Hattori M, Parkhill J, Sebaihia M, Coulthurst SJ, Gotoh N, Thomson NR, Ewbank JJ, Hayashi T (2014) Genome evolution and plasticity of Serratia marcescens, an important multidrug-resistant nosocomial pathogen. Genome Biol Evol 6(8):2096–2110. https://doi.org/10.1093/gbe/evu160
Jarostawiecka A, Pietrowska-Seget Z (2014) Lead resistance in micro-organisms. Microbiology 160(1):12–25. https://doi.org/10.1099/mic.0.070284-0
Jorquera MA, Hernández M, Martínez O, Marschner P, De M, Mora L (2010) Detection of aluminium tolerance plasmids and microbial diversity in the rhizosphere of plants grown in acidic volcanic soil. Eur J Soil Biol 46(3–4):255–263. https://doi.org/10.1016/j.ejsobi.2010.03.005
Júnior RAB, Rivera FP, Moreira J (2016) A practical guide to biospeckle laser analysis—theory and software. Editora UFLA, p 158. ISBN: 9-788581-270517
Kästner M, Breuer-Jammali M, Mahro B (1994) Enumeration and characterization of the soil microflora from hydrocarbon-contaminated soil sites able to mineralize polycyclic aromatic hydrocarbons (PAH). Appl Microbiol Biotechnol 41:267–273. https://doi.org/10.1007/BF00186971
Khan AR, Park GS, Asaf S, Hong SJ, Jung BK, Hin JH (2017) Complete genome analysis of Serratia marcescens RSC-14: a plant growth-promoting bacterium that alleviates cadmium stress in host plants. PLoS ONE 12:1–17. https://doi.org/10.1371/journal.pone.0171534
Kiran M, Pakshirajan K, Das G (2017) Heavy metal removal from multicomponent system by sulfate reducing bacteria: Mechanisms and cell surface characterization. J Hazard Mater 324:62–70. https://doi.org/10.1016/j.jhazmat.2015.12.042
Kotoky R, Pandey P (2020) Rhizosphere assisted biodegradation of benzo(a)pyrene by cadmium resistant plant-probiotic Serratia marcescens S2I7, and its genomic traits. Sci Rep 10:5279. https://doi.org/10.1038/s41598-020-62285-4
Kotoky R, Nath S, Mahes-hwari DK, Pandey P (2019) Cadmium resistant plant growth promoting rhizobacteria Serratia marcescens S2I7 associated with the growth promotion of rice plant. Environ Sustain 2:135–144. https://doi.org/10.1007/s42398-019-00055-3
Kumar R, Acharya C, Joshi SR (2011) Isolation and analyses of uranium tolerant Serratia marcescens strains and their utilization for aerobic uranium U(VI) bioadsorption. J Microbiol 49(4):568–574. https://doi.org/10.1007/s12275-011-0366-0
Limcharoensuk T, Sooksawat N, Sumarnrot A, Awutpet T, Kruatrachue M, Pokethitiyook P, Auesukaree C (2015) Bioaccumulation and biosorption of Cd2+and Zn2+ by bacteria isolated from a zinc mine in Thailand. Ecotoxicol Environ Saf 122:322–330. https://doi.org/10.1016/j.ecoenv.2015.08.013
Lemos LN, Medeiros JD, Andreote FD, Fernandes GR, Varani AM, Oliveira G, Pylro VS (2019) Genomic signatures and co-occurrence patterns of the ultra-small Saccharimonadia (phylum CPR/Patescibacteria) suggest a symbiotic lifestyle. Mol Ecol 29(10):1936–1936. https://doi.org/10.1111/mec.15208
Leng F, Li Y, Luo W, Wei Q, Jing Y, Wang X, Yang M, Wang Y (2019) Cloning, expression, and bioinformatics analysis of heavy metal resistance gene afe_1862 from Acidothiobacillus ferroxidans L1 in Escherichia coli. Biol Trace Elem Res 189:291–300. https://doi.org/10.1007/s12011-018-1462-3
Lu M, Li Z, Liang J, Wei Y, Rensing C, Wei G (2016) Zinc resistance mechanisms of P1B-type ATPases in Sinorhizobium meliloti CCNWSX0020. Sci Rep 6:29355. https://doi.org/10.1038/srep29355
Marrero J, Auling G, Coto O, Nies DH (2007) High-level resistance to cobalt and nickel but probably no transenvelope efflux: metal resistance in the Cuban Serratia marcescens strain C-1. Microb Ecol 53:123–133. https://doi.org/10.1007/s00248-006-9152-7
MATLAB (2010) The language of technical computing. version 7.10.0 (R2010a), The MathWorks Inc., Natick. https://www.mathworks.com/?s_tid=gn_logo. Accessed 10 Jun 2020
Mehrotra A, Kundu K, Sreekrishnan TR (2016) Decontamination of heavy metal laden sewage sludge with simultaneous solids reduction using thermophilic sulfur and ferrous oxidizing species. J Environ Manag 167:228–235. https://doi.org/10.1016/j.jenvman.2015.11.004
Munoz AJ, Ruiz E, Abriouel H, Galvez A, Ezzouhri L, Lairini K, Espínola F (2012) Heavy metals tolerance of microorganisms isolated from wastewaters: identification and evaluation of its potential for biosorption. Chem Eng J 210:325–332. https://doi.org/10.1016/j.cej.2012.09.007
Naik MM, Dubey SK (2013) Lead resistant bacteria: Lead resistance mechanisms, their applications in lead bioremediation and biomonitoring. Ecotoxicol Environ Saf 98:1–7. https://doi.org/10.1016/j.ecoenv.2013.09.039
Newsome L, Morris K, Lloyd JR (2015) Uranium biominerals precipitated by an environmental isolate of Serratia under anaerobic conditions. PLoS ONE 10(7):1–14. https://doi.org/10.1371/journal.pone.0132392
Nies DH (2003) Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 27(2–3):313–339. https://doi.org/10.1016/S0168-6445(03)00048-2
Nong Q, Yuan K, Li Z, Chen P, Huang Y, Hu L, Jiang J, Luan T, Chen B (2019) Bacterial resistance to lead: chemical basis and environmental relevance. J Environ Sci 85:46–55. https://doi.org/10.1016/j.jes.2019.04.022
Permina EA, Kazakov AE, Kalinina OV, Gelfand MS (2006) Comparative genomics of regulation of heavy metal resistance in Eubacteria. BMC Microbiol 6:1–11. https://doi.org/10.1186/1471-2180-6-49
Pires JF, Cardoso LD, Schwan RF, Silva CF (2017) Diversity of microbiota found in coffee processing wastewater treatment plant. World J Microbiol Biotechnol 33(12):1–12. https://doi.org/10.1007/s11274-017-2372-9
Pires JF, Viana DC, Braga RA, Schwan RF, Silva CF (2021) Protocol to select efficient microorganisms to treat coffee wastewater. J Environ Mana 278:111541. https://doi.org/10.1016/j.jenvman.2020.111541
Retchless AC, Lawrence JG (2007) Temporal fragmentation of speciation in bacteria. Science 317(5841):1093–1096. https://doi.org/10.1126/SCIENCE.1144876
Sheoran AS, Sheoran V (2006) Heavy metal removal mechanism of acid mine drainage in wetlands: a critical review. Miner Eng 19(2):105–116. https://doi.org/10.1016/j.mineng.2005.08.006
Shukor MY, Habib SHM, Rahman MFA, Jirangon H, Abdullah MPA, Shamaan NA, Syed M (2008) Hexavalent molybdenum reduction to molybdenum blue by S. Marcescens strain Dr. Y6. Appl Biochem Biotechnol 149(1):33–43. https://doi.org/10.1007/s12010-008-8137-z
Taghavi S, Lesaulnier C, Monchy S, Wattiez R, Mergeay M, Lelie D (2009) Lead(II) resistance in Cupriavidus metallidurans CH34: interplay between plasmid and chromosomally-located functions. Antonie Van Leeuwenhoek 96:171–182. https://doi.org/10.1007/s10482-008-9289-0
Thompson AS, Maani EV, Lindell AH, King CJ, McArthur JV (2007) Novel Tetracycline Resistance Determinant Isolated from an Environmental Strain of Serratia marcescens. Appl Environ Microbiol 73(7):2199–2296. https://doi.org/10.1128/AEM.02511-06
Viana DC, Pires JF, Braga R (2017) Biospeckle laser technique applied for estimating disinfection accomplishment of wastewaters subjected to chlorination. Process Saf Environ Protection 109:670–676. https://doi.org/10.1016/j.psep.2017.05.017
Vijayaraghavan K, Yun YS (2008) Bacterial biosorbents and biosorption. Biotechnol Adv 26(3):266–291. https://doi.org/10.1016/j.biotechadv.2008.02.002
Wang T, Chen K, Gao F, Kang Y, Chaudhry MT, Wang Z, Wang Y, Shen X (2017) ZntR positively regulates T6SS4 expression in Yersinia pseudotuberculosis. J Microbiol 55:448–456. https://doi.org/10.1007/s12275-017-6540-2
WHO (2017) Guidelines for drinking-water quality: fourth edition incorporating the first addendum. World Health Organization, Geneva, p 631
Acknowledgements
The authors thank Brazilian agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico do Brasil (CNPQ), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for scholarship, as well as Centro de Ciências do Mar (CCMAR) through projects UIDB/04326/2020), (UIDP/04326/2020) and (LA/P/0101/2020) and FCT co-financed by the Algarve´s Regional Operational Program (CRESC Algarve 2020) through Portugal 2020 and European Regional Development Fund (FEDER), under the project METALCHEMBIO (no. 29251).
Funding
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Centro de Ciências do Mar (CCMAR) (UIDB/04326/2020), (UIDP/04326/2020) and (LA/P/0101/2020). Funds from FCT co-financed by the Algarve´s Regional Operational Program (CRESC Algarve 2020) through Portugal 2020 and European Regional Development Fund (FEDER), under the project METALCHEMBIO (no. 29251).
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. The first draft of the manuscript was written, reviewed, and edited by GMRF that was also responsible for material preparation, methodology development, data collection, validation data and formal data evaluation. All authors commented on previous versions of the manuscript. JFP was responsible for material preparation, methodology development, data collection, validation data and formal data evaluation. All authors commented on previous versions of the manuscript. LSR made critical, specific comments, reviewed the presentation of the data, and edited the manuscript. JDC supervised Gustavo Ferreira in the search for gene sequences in world databases, the design of primers, the optimization of PCR conditions, the analysis and selective trimming of DNA sequences and their alignment and contigs construction, as well as in the search of Open Reading Frames (ORFs). MCC supervised Gustavo Ferreira and made critical, specific comments, reviewed the presentation of the data, and edited the manuscript. RFS made critical, specific comments, reviewed the presentation of the data, and edited the manuscript. CFS supervised Gustavo Ferreira in whole experiments period in Brazil, also performed the original writing of the manuscript, supervision, and project administration. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors have no conflicts of interest to declare that are relevant to the content of this article.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
dos Reis Ferreira, G.M., Pires, J.F., Ribeiro, L.S. et al. Impact of lead (Pb2+) on the growth and biological activity of Serratia marcescens selected for wastewater treatment and identification of its zntR gene—a metal efflux regulator. World J Microbiol Biotechnol 39, 91 (2023). https://doi.org/10.1007/s11274-023-03535-1
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-023-03535-1