Chromium bioremediation by Alcaligenes sp. strain newly isolated from chromite mine of Sabzevar

Document Type : Original Article


1 Department of Biology, Faculty of Basic Sciences, Hakim Sabzevari University, Sabzevar, Iran

2 Department of Biochemistry and Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran


In this work, CKCr-6A strain was found to be highly resistant to some toxic heavy metals such as Cr+6, Cr+3, Cu+2, Co+2, Cd+2, Pb+2, Hg+2, U+6, tellurium, and selenite. Herein, high chromate tolerance of an isolated strain is reported with a high minimum inhibitory concentration value of 80,000 mg/L and the effective parameters (pH, temperature, shaking, and glucose concentration) were selected for Cr(VI) removal by this isolated strain. Cr(VI) elimination by the target strain increased with glucose addition to the culture medium. We optimized the possible parameters and their interactions using design experimental software. After optimization, this strain showed high efficiency in detoxifying chromate; this could reduce up to 100 mg/L of Cr(VI) to Cr(III) over 3 h. The CKCr-6A strain exhibited ability to in vitro reduction after 3 h and repeated removing of Cr(VI) without any amendment of nutrients, suggesting its possible application in continuous bioremediation.


  1. Chuan MC, Liu JC. Release behavior of chromium from tannery sludge. Water Res 1996; 30: 932-938.
  2. Basu M, Bhattacharya S, Paul AK. Isolation and characterization of chromium-resistant bacteria from tannery effluents. Bull Environ Conta Toxicol 1997; 58: 535–542.
  3. Caravelli A.H, Giannuzzi L, Zaritzky NE. Reduction of hexavalent chromium by Sphaerotilus natans a filamentous micro-organism present in activated sludges. J Hazard Mater 2008; 156: 214–222.
  4. O'Brien TJ, Ceryak S,  Patierno SR. Complexities of chromium carcinogenesis: role of cellular response, repair and recovery mechanisms. Mutat Res 2003; 533: 3-36.
  5. Beyersmann D, Hartwig A. Carcinogenic metal compounds: recent insight into molecular and cellular mechanisms. Arch Toxicol 2008; 82: 493-512.
  6. Park D, Yun YS, Park JM. Studies on hexavalent chromium biosorption by chemically-treated biomass of Ecklonia sp. Chemosphere 2005; 60: 1356–1364.
  7. Mungasavalli DP, Viraraghavan T,  Jin YC. Biosorption of chromium from aqueous solutions by pretreated Aspergillus niger: Batch and column studies. Colloid Surf Phys Chem Eng Aspects 2007; 301: 214–223.
  8. Quintelas C, Fernandes B, Castro J, Figueiredo H,  Tavares T. Biosorption of Cr(VI) by a Bacillus coagulans biofilm supported on granular activated carbon (GAC). Chem Eng J 2008; 136: 195-203.
  9. Frankel RB, Bazylinski DA. Biologically Induced Mineralization by Bacteria. Rev Miner Geochem 2003; 54: 95-114.
  10. Rohwerder T, Gehrke T, Kinzler K, Sand W. Bioleaching review part A: progress in bioleaching: fundamentals and mechanisms of bacterial metal sulfide oxidation. Appl Microbiol Biotechnol 2003; 63: 239–48.
  11. Elangovan R, Abhipsa S, Rohit B, Ligy P, Chandraraj K. Reduction of Cr(VI) by a Bacillus sp. Biotechnol Lett 2006; 28: 247–252.
  12. Kiran B, Kaushik A,  Kaushik CP. Biosorption of Cr(VI) by native isolate of Lyngbya putealis (HH-15) in the presence of salts. J Hazard Mater 2007; 141: 662–667.
  13. Kalil SJ, Maugeri F,  Rodrigues MI. Response surface analysis and simulation as a tool for bioprocess design and optimization. Proc Biochem 2000; 35: 539- 550.
  14. Myers RH, Montgomery DC, Anderson-Cook C. Response Surface Methodology: Process and Product Optimization Using Designed Experiments. Wiley, New York 2002.
  15. Mollania N, Mesbahi-Norozi M, Novrozi-Nejad Z, Mollania H,  Khajeh K. High Ability of Bacillus Cereus Strain Isolated from Chromite Mine in Bioremediation of Carcinogenic Chromium-Contaminated Wastewater. Casp J Appl Sci Res 2013;  2: 10-12.
  16. Mollania N, Tayebee R, Narenji-Sani F. An environmentally benign method for the biosynthesis of stable selenium nanoparticles. Res Chem Intermed 2016; 42: 4253-4271.
  17. Mollania N, Mesbahi- Nowrouzi M. Purification of selenate reductase from Alcaligenes sp. CKCr-6A with the ability to biosynthesis of selenium nanoparticle: Enzymatic behavior study in imidazolium based ionic liquids and organic solvent. J Mol Liq 2018; 249: 1254–1262.
  18. Bauernfeind A. Antibiotic susceptibility patterns of respiratory isolates of Klebsiella pneumoniae in Europe and the USA in 1992 and 1993, The Alexander Project Collaborative Group. J Antimicrob Chemother 1996; 38: 107-115.
  19. Collee JG, Faster AG, Maromion BP, Simmons A (eds), Mackie & McCartney. Practical Medical Microbiology. Published by Churchill Livingstone (1996).
  20. Sambrook J, Russell DW. Molecular Cloning a Laboratory Manual. Cold Spring Harbor, NewYork 2001.
  21. DeSantis TZ, Dubosarskiy I, Murray S, Andersen GL. Comprehensive aligned sequence construction for automated design of effective probes (CASCADE-P) using 16S rDNA. Bioinformatics 2003; 19: 1461–1468.
  22. Rice EW, Baird RB, Eaton AD, Clesceri LS. Standard Methods for the Examination of Water and Wastewater Washington, DC, USA 1981.
  23. Yan JY, Yan YJ. Optimization for producing cell-bound lipase from Geotrichum sp. and synthesis of methyl oleate in microaqueous solvent. Appl Microbiol Biotechnol 2008; 78: 431- 439.
  24. Viti C, Pace A, Giovannetti L. Characterization of Cr(VI)-resistant bacteria isolated from chromium-contaminated soil by tannery activity. Curr Microbiol 2003; 46: 1-5.
  25. Ibrahim ASS, El-Tayeb MA, Elbadawi YB,  Al-Salamah AA. Bioreduction of Cr (VI) by potent novel chromate resistant alkaliphilic Bacillus sp. strain KSUCr5 isolated from hypersaline Soda lakes. Afr J Biotechnol 2011; 37: 7207-7218.
  26. Calomiris JJ, Armstrong JL, Seidler RJ. Association of metal tolerance with multiple antibiotic resistance of bacteria isolated from drinking water. Appl Environ Microbiol 1984; 47: 1238–1242.
  27. Verma T, Srinath T, Gadpayle RU, Ramteke PW, Hans RK, Garg SK. Chromate tolerant bacteria isolated from tannery effluent. Bioresour Technol 2001; 78: 31-35.
  28. Silver S,  Misra TK. Plasmid-mediated heavy metal resistances. Annu Rev Microbiol 1988; 42: 717-743.
  29. Cheung K, Lai H, Gu J. Membrane-associated hexavalent chromium reductase of Bacillus megaterium TKW3 with induced expression. J Microbiol Biotech 2006; 16: 855–862.
  30. Bae WC, Lee HK, Choe YC, Jahng DJ, Lee SH, Kim SJ. Purification and characterization of NADPH-dependent Cr(VI) reductase from Escherichia coli ATCC 33456. J Microbiol 2005; 43: 21–27.
  31. Desai C, Jain K, Madamwar D. Evaluation of in vitro Cr(VI) reduction potential in cytosolic extracts of three indigenous Bacillus sp. isolated from Cr(VI) polluted industrial landfill. Bioresour Technol 2008; 99; 6059–6069.
  32. McLean J,  Beveridge TJ. Chromate reduction by a pseudomonad isolated from a site contaminated with chromated copper arsenate. Appl Environ Microbiol 2001; 67: 1076–1084.
  33. Pal A, Dutta S,  Paul AK. Reduction of hexavalent chromium by cell-free extract of Bacillus sphaericus AND 303 isolated from serpentine soil. Curr Microbiol 2005; 51: 327–330.
  34. Park CH, Keyhan M, Wielinga B, Fendorf S,  Matin A. Purification to Homogeneity and Characterization of a Novel Pseudomonas putida Chromate Reductase. Appl Environ Microbiol 2000; 66: 1788–1795.
  35. Li ZY, Guo SY,  Li L. Study on the process, thermodynamical isotherm and mechanism of Cr(III) uptake by Spirulina platensis. J Food Eng 2006; 75; 129–136.
  36. Peitzsch N, Eberz G, Nies DH. Alcaligenes eutrophus as a bacterial chromate sensor. Appl Environ Microb 1998; 64: 453-458.
  37. Camargo FA, Bento FM, Okeke BC, Frankenberger WT. Chromate reduction by chromium-resistant bacteria isolated from soils contaminated with dichromate. J Environ Qual 2003; 32: 1228–1233.
  38. Faisal M, Hasnain S. Microbial conversion of Cr (VI) in to Cr (III) in industrial effluent. Afr J Biotechnol 2004; 3; 610-617.
  39. Dhal B, Thatoi H,. Das N, Pandey BD. Reduction of hexavalent chromium by Bacillus sp. isolated from chromite mine soils and characterization of reduced product. J Chem Technol Biotechnol 2010; 85: 1471–1479.
  40. Lawson JS, Erjavec J. Modern Statistics for Engineering and Quality Improvement. Brooks/Cole-Thomson Learning: Pacific Grov 2000.
  41. Lameiras S, Quintelas C, Tavares T. Biosorption of Cr (VI) using a bacterial biofilm supported on granular activated carbon and on zeolite. Bioresour Technol 2008; 99; 801–806.
  42. Pandi M, Shashirekha V, Swamy M. Bioabsorption of chromium from retan chrome liquor by cyanobacteria. Microbiol Res 2009; 164: 420-428.
  43. Doshi H, Ray, A, Kothari IL. Biosorption of cadmium by live and dead Spirulina: IR spectroscopic, kinetics, and SEM studies. Curr Microbiol 2007; 54: 213–218.
  44. Das SK, Guha AK. Biosorption of chromium by Termitomyces clypeatus. Colloids Surf BBiointerfraces 2007; 60: 46–54.
  45. Aravindhan R, Madhan B, Rao JR, Nair BU, Ramasami T. Bioaccumulation of chromium from tannery wastewater: an approach for chrome recovery and reuse. Environ Sci Technol 2004; 38: 300–306.
  46. Loukidou MX, Zouboulis AI, Karapantsios TD,  Matis KA. Equilibrium and kinetic modeling of chromium (VI) biosorption by Aeromonas caviae. Colloids Surf Phys Chem Eng Aspects 2004; 242: 93–104.
  47. Ferraro JR. Infrared spectra of inorganic and coordination compounds (Nakamoto, Kazuo). J. Chem. Educ 1963; 40 (9):501.