Arsenic Bioremediation Potential of Arsenite Oxidizing Bacteria Isolated from Geogenic and Anthropogenically Contaminated Soil

Document Type : Original Research Paper


Department of Environmental and Life Sciences, IIS (deemed to be) University, SFS, Mansarovar, 302020 Jaipur (Rajasthan), India


The soil of many places of eastern India contains high amount of arsenic, due to several geogenic activities in this area. In the specific regions of the country where there is no such type of Geogenic activities, the soil is found to be almost free of arsenic. In such places where there are industries, the soil is being contaminated with the arsenic due to anthropogenic activities. One of such site which was selected for the study was in close vicinity to the textile industries in Jaipur, Rajasthan, India discharging their effluents having 423 µg/g arsenic. While the soil sample collected from the far eastern part of Tezpur Assam, India, contaminated by Geogenic sources contained 443µg/g arsenic. Four arsenite resistant bacterial strains were isolated from each of the samples. Strains SE-3 and TB-1 isolated from Jaipur and Tezpur, respectively showed highest minimum inhibitory concentration of 46.5mM and 38.7mM sodium arsenite. Based on 16S rDNA sequencing and nucleotide homology and Phylogenetics analysis strain, SE-3 was identified as Pseudomonas sp. SE-3 (accession no. KP730605) and TB-1 as Bacterium TB-1 (accession no KP866680). Complete oxidation of arsenite to less toxic form arsenate was observed in Pseudomonas sp. SE-3, while 64.6% by Bacterium TB-1. The arsenite oxidation was supported on the molecular level by confirming the presence of aox gene by PCR amplification. The enzyme activity of arsenite oxidase was also established. Arsenic hyper tolerant bacteria isolated from these soils having arsenite oxidizing ability show a promising way for the bioremediation of arsenic in contaminated soil.


Abbas, S. Z., Riaz, M., Ramzan, N., Zahid, M. T., Shakoori, F. R. and Rafatullah, M. (2015). Isolation and characterization of arsenic resistant bacteria from wastewater. Brazilian journal of microbiology : publication of the Brazilian Society for Microbiology., 45(4); 1309–1315.
Aksornchu, P., Prasertsan, P. and Sobhon V. (2008).Isolation of arsenic-tolerant bacteria from arsenic-contaminated soil. Songklanakarin, J. Sci. Technol., 30; 95-102.
Anderson, G.L., Williams, J. and Hille, R. (1992).The purification and characterization of arsenite oxidase from Alcaligenes faecalis, a molybdenum containing hydroxylase.J. Biol. Chem., 267; 23674-23682.
Bachate, S.P., Khapare, R.M. and Kodam, K.M. (2012). Oxidation of arsenite by two betaproteobacteria isolated from soil. Appl. Microbiol. Biotechnol., 93; 2135–2145.
Bora, P.K., Chetry, S., Sharma, D.k. and Saikia, P. M. (2013). Distribution Pattern of Some Heavy Metals in the Soil of Silghat Region of Assam (India), Influenced by Jute Mill Solid Waste. J.Chem., Article ID 609203, 7 pages.
Cai, L., Liu, G., Rensing, C. and Wang, G. (2009a). Genes involved in arsenic transformation and resistance associated with different levels of arsenic contaminated soils. BMC Microbiol., 9; 4.
Campos, V. L.,  León,C., Mondaca, M. A., Yañez, J. and  Zaror, C. (2010). Arsenic Mobilization by Epilithic Bacterial Communities Associated with Volcanic Rocks from Camarones River, Atacama Desert, Northern Chile, Arch. Environ. Contam. Toxicol., 61(2); 185-192.
Chang, J.S., Yoon, I.H., Lee, J.H., Kim, K.R., An, J. and Kim, K.W. (2010). Arsenic detoxification potential of aox genes in arsenite-oxidizing bacteria isolated from natural and constructed wetlands in the Republic of Korea. Enviro. Geochem. Health., 32; 95-105.
Chaturvedi, N. and Pandey, P. N. (2014).Phylogenetic Analysis of Gammaproteobacterial Arsenate Reductase Proteins Specific to Enterobacteriaceae Family, Signifying Arsenic Toxicity. Interdiscip Sci Comput Life Sci., 6;1-6.
Chiroma, T.M., Ebewele, R.O. and Hymore, F.K. (2014). Comparative assessment of heavy metal levels in soil, vegetables and urban grey waste water used for irrigation in Yola and Kano. Int. refereed j. eng. sci., 3; 01-09.
Dave, D., Sarma, S., Parmar, P., Shukla, A., Goswami, D., Shukla, A. and Saraf, M. (2020). Microbes as a boon for the bane of heavy metals. Environmental Sustainability. 3(3); 233-255 doi:10.1007/s42398-020-00112-2. 
Derome, J. (2000). Detoxification and amelioration of heavy metal contaminated forest soils by means of liming and fertilization. Environ. Pollut., 17;79-88. 
Deshpande, L.S. and Pande, S.P. (2005). Development of arsenic testing field kit -a tool for rapid on-site screening of arsenic contaminated water sources. Environ. Monit. Assess., 101(1); 93-101. 
Dey, U., Chatterjee, S, and Mondal, N. (2016).Isolation and characterization of arsenic-resistant bacteria and possible application in bioremediation. Biotechnol. Rep., 10; 1-7.
Diliana, D., Simeonova.,Lièvremont, D., Lagarde,F., Daniel A., Muller, E., Veneta I., Groudev. and Marie-Claire (2004).Microplate screening assay for the detection of arsenite-oxidizing and arsenate reducing bacteria. FEMS Microbiol.Lett., 249-253.
Durve, A., Naphade, S., Bhot, M., Varghese, J. and Chandra., N. (2012).Characterisation of metal and xenobiotic resistance in bacteria isolated from textile effluent. Adv. Appl. Sci. Res., 3(5); 2801-2806.
Ghosh, S. and Sar, P. (2013). Identification and characterization of metabolic properties of bacterial populations recovered from arsenic contaminated ground water of North East India (Assam).Water Res., 47(19); 6992-7005. 
Ghosh, D., Bhadury, P. and Routh, J. (2014).Diversity of arsenite oxidizing bacterial communities in arsenic-rich deltaic aquifers in West Bengal, India.Front Microbiol., 5; 602.
Gikas, P., Sengor, S., Ginn, T., Moberly, J. and Peyton, B. (2009).The effects of heavy metals and temperature on microbial growth and lag.Global NEST J Journal., 11;325–332.
Green, H.H. (1918). Description of a bacterium which oxidizes arsenite to arsenate, and of one which reduces arsenate to arsenite, isolated from a cattle dipping tank. S. Afr. J. Sci., 14; 465-467.
Hoeft, S. E., Blum, J. S., Stolz, J. F., Tabita, F. R., Witte, B., King, G. M., Santini, J. M. and Oremland, R.S. (2007). Alkalilimnicolaehrlichii sp. nov., a novel, arsenite-oxidizing haloalkaliphilic gamma proteobacterium capable of chemo autotrophic or heterotrophic growth with nitrate or oxygen as the electron acceptor. Int. J. Syst. Evol. Microbiol., 57; 504–512.
Hu, S., Lu, J. and Jing, C. (2012).A novel colorimetric method for field arsenic speciation analysis. J Environ Sci., 24 (7); 1341–1346.
Joshi, D. N.,  Patel, J. S.,  Flora, S. J. S. and  Kalia, K. (2008).Arsenic accumulation by Pseudomonas stutzeri and its response to some thiol chelators, Environ Health Prev Med., 13(5); 257–263.
Kim, S.J. (1985) Effects of heavy metals on natural populations of bacteria from surface micro-layers and sub-surface waters. Mar. Ecol. Prog. Ser., 26; 203-206.
Koechler, S., Cleiss-Arnold, J., Proux, C., Sismeiro, O., Dillies, M.A., Goulhen-Chollet, F., Hommais, F., Lievremont, D., Arsene-Ploetze, F., Coppee, J.Y. and Bertin, P.N. (2010). Multiple controls affectsarsenite oxidase gene expression in Herminiimonas arsenic oxydans. BMC Microbiol.,10; 53.
Laverman, A.M., Switzer, B. J., Schaefer, J.K., Philips, E.J.P., Lovley, D.R. and Oremland, R.S. (1995).Growth of strain SES-3 with arsenate and other diverse electron acceptors.Appl Environ Microbiol., 61; 3556–3561.
Li, X., Gong, J., Hu, Y., Cai, L., Johnstone, L., Grass, G., Rensing, C. and Wang, G. (2012).Genome sequence of the moderately halotolerant, arsenite-oxidizing bacterium Pseudomonas stutzeri TS44. J. Bacteriol., 194(16); 4473-4.
Lieutaud, A., Lis, R.V., Duval, S., Capowiez, L., Muller, D.,Lebrun, R.,Lignon, S., Fardeau, M.L., Lett, M.C., Nitschke, W. and Cothenet, B.C. (2010).Arsenite Oxidase From Ralstonia Sp. 22Characterization Of The Enzyme and Its Interaction With Soluble Cytochromes. J Biol Chem., 285(27); 20433–20441.
Mahawar,P. and Akhtar, A. (2015). Physico-Chemical Characterization of Soil and Effluent of Dye Industries in Kaithun region of Kota, Rajasthan. Int. J. Pure App. Biosci., 3 (2); 419-422.
Maiti, S.K. (2003). Analysis of Physical Parameters of Soil (Chapter 10) in book: Handbook of Methods in Environmental Studies: Air, Noise, Soil and Overburden analysis. ABD Publishers, Jaipur, India., 2; 142-161.
Majumder, A. (2012). Arsenic Resistant Bacteria Isolated From Contaminated Soil And Selection Of Arsenic Reducing Strains. The Bioscan., 7(3); 469-472.
Mandal, B.K. and Suzuki, KT (2002). Arsenic round the world: a review. Talanta., 58(1); 201-235.
Mobar, S., Kaushik, P. and Bhatnagar, P. (2015). Physiochemical Comparison of Textile Effluent Impacted And Un- Impacted Agricultural Soil Of Jaipur City, India. Int. J. Recent Sci. Res., 6(3); 3090-3093.
Muehe, E.M. and Kappler, A. (2014).Arsenic mobility and toxicity in South and South-east Asia – a review on biogeochemistry, health and socio-economic effects, remediation and risk predictions. Environ. Chem., 11; 483–495.
Muller, D., Lièvremont, D., Simeonova, D.D., Hubert, J.C. and Lett, M.C. (2003).Arsenite oxidase aox genes from a metal-resistant beta-proteobacterium. J Bacteriol.,185; 135-141.
Nakagawa,T.,   Iino, T.,   Suzuki, K. and  Harayama, S. (2006). Ferrimonas futtsuensis sp. nov. and Ferrimonas kyonanensis sp. nov., selenate-reducing bacteria belonging to the Gamma proteobacteria isolated from Tokyo Bay. Int. J. Syst. Evol. Microbiol., 56; 2639-2645.
Pegu, B.K, Buragohain, N., Nakoti, N., Sarmah, P. and Das, S. (2019). Isolation and characterization of arsenic resistant Bacteria from groundwater from lakhimpur district. Asian. J. Microbiol. Biotechnol. Environ. Sci., 21; S94-S100.
Peterson, R. and Jensén, P. (1989).The role of bacteria in pH increase of nettle water. Plant Soil., 113; 137–140.
Prasad, K.S., Subramanian, V. and Paul, J. (2009).Purification and characterization of arsenite oxidase from Arthrobacter sp. Biometals., 22; 711-721.
Pratush, A., Kumar, A. and Hu, Z. (2018). Adverse effect of heavy metals (As, Pb, Hg, and Cr) on health and their bioremediation strategies: a review. Int. Microbiol., 21(3); 97-106. doi:10.1007/s10123-018-0012-3.
Quéméneur, M., Cébron, A., Billard, P., Battaglia-Brunet, F., Garrido, F., Leyval, C. and Joulian, C. (2010). Population structure and abundance of arsenite-oxidizing bacteria along an arsenic pollution gradient in waters of the upper isle River Basin, France. Appl.Environ.Microbiol., 76(13);4566–
Rahaman, S., Sinha, A., C and Pati, R. (2013). Arsenic contamination: a potential hazard to the affected areas of West Bengal, India. Environ. Geochem. Health., 35; 119-132.  
Sabhapandit, P., Saikia,  P. and Mishra, A.K. (2010). Statistical analysis of heavy metals from water samples of tezpur sub-division in Sonitpur district, Assam, India. Int. J. Appl. Biol. Pharm., 1(3); 946-951.
Sahajrao, N.M. and Pawale, R.G. (2014). Characteristics Of Soil & Contamination Of Heavy Metals From The Catchment Area Manjara River In BiloliTaluka, Nanded, Maharashtra. Scientia Research Library, Journal of Applied Science And Research., 2(6); 78-85.
Salmeron, H.A.,   Cordi, A., Armanet, C. B.,  Halter, D., Pagnout, C., fard, E. A.,   Montaut, D.,  Seby, F.,  Bertin, P. N.,  Bauda, P. and  Ploetze, F. A. (2011). Unsuspected Diversity of Arsenite-Oxidizing Bacteria as Revealed by Widespread Distribution of the aoxB Gene in Prokaryotes. Appl. Environ. Microbiol., 77(13); 4685–4692.
Selvi, M. S., Sasikumar, S., Gomathi, S.,Rajkumar, P., Sasikumar, P. and Sadasivam, S. G. (2014).Isolation and characterization of arsenic resistant bacteria from agricultural soil, and their potential for arsenic bioremediation. International Journal of Agricultural Policy and Research., 2(11); 393-405.
Shakya, S. and Pradhan, B. (2013). Isolation and characterization of arsenic resistant pseudomonas stutzeri asp3 for its potential in arsenic resistance and removal. Kathmandu University.  J. Eng. Sci. Technol., 9(1); 48-59.
Sharma, B., Singh, S. and Siddiqi, N.J. (2014). Biomedical Implications of Heavy Metals Induced Imbalances in Redox Systems. BioMed. Res. Int., 640-754.
Sher, S. and Rehman, A. (2019).Use of heavy metals resistant bacteria-a strategy for arsenic bioremediation.Appl. Microbiol. Biotechnol., 103(15); 6007-6021. doi: 10.1007/s00253-019-09933-6. 
Simeonova, D.D., Lievremont, D., Lagarde, F., Muller, D.A.E., Groudeva, V.L. and Lett, M-C. (2004). Microplate screening assay for the detection of arsenite oxidizing and arsenate reducing bacteria. FEMS Microbiol.Lett., 237; 249-253.
Standard methods for examination of water and waste water-APHA (2005); Part 2000, 3000, 3500. American Public Health Association (APHA)-American water works Association and Water Environment Federation. Centennial Edition.
Sun, W., Sierra, R. and Field, J.A. (2008). Anoxic oxidation of arsenite linked to denitrification in sludges and sediments. Water Res., 42; 4569–4577. 
Taran, M., Fateh, R., Rezaei, S. and Gholi, M. K. (2019).Isolation of arsenic accumulating bacteria from garbage leachates for possible application in bioremediation. Iran. J.Microbiol., 11(1); 60–66.
Wan, X., Lei, M. and Chen, T. (2019). Review on remediation technologies for arsenic-contaminated soil.  Front. Environ. Sci. Eng., 14(2). doi:10.1007/s11783-019-1203-7.