Cr(VI) mediated hydrolysis of algae cell walls to release TOC for enhanced biotransformation of Cr(VI) by a culture of Cr(VI) reducing bacteria

  • M. M. RoestorffEmail author
  • E. M. N. Chirwa


Hexavalent chromium [Cr(VI)], the most toxic form of chromium, is frequently released into the environment from anthropogenic sources. Cr(VI) mainly occurs in the oxyanionic forms, CrO42− and Cr2O72−. It is highly oxidative and carcinogenic under chronic and subchronic exposure conditions. Conventionally, Cr(VI) pollution is remediated by reducing Cr(VI) to Cr(IIII). Cr(III) is naturally less toxic than Cr(VI) and is a 1000 times less mobile in the aquatic phase than Cr(VI). Biological reduction and detoxification of Cr(VI) are viewed as the most ecologically friendly process for remediation of Cr(VI) pollution. However, fast reduction of Cr(VI) mainly occurs under aerobic conditions in the presence of organic carbon sources. In the current research, freshwater algae are utilized as a carbon source for Cr(VI) reduction with using symbiotic bacterial cultures. The algal species, Chlamydomonas reinhardtii and Chlorococcum ellipsoideum, were tested in their ability to serve as or produce a carbon source for locally isolated bacteria to achieve reduction of Cr(VI) to Cr(III). Batch experiments were conducted under aerobic conditions at different concentrations of Cr(VI) to determine the kinetics of the biological reduction reaction. In the batch experiments, complete removal of up to 50 mg L−1 of initial Cr(VI) concentration was achieved within 24 h. At 100 mg L−1 initial Cr(VI) concentration, the system could remove 92% of the Cr(VI). Algae was found to be very sensitive to Cr(VI) toxicity. The Cr(VI) inhibited the algae growth and reduced the chlorophyll a content and by extension the algae’s ability to undergo photosynthesis.


Phytoremediation Freshwater algae Cr(VI) reduction Bioremediation Batch kinetic 


Funding information

The authors would like to thank the University of Pretoria and the Water Utilization and Environmental Engineering Division at the University of Pretoria for the research support. The research was partially funded by the National Research Foundation (NRF) of South Africa through Grant No. CSUR180215313534 awarded to Prof Evans M. N. Chirwa of the Department of Chemical Engineering and Maria Roestorff (Grant No: 114172). Additional funding was provided by the Sedibeng Water Chair in Water Utilization Engineering in the Water Utilization Division at the University of Pretoria.

Supplementary material

10811_2018_1716_MOESM1_ESM.docx (67 kb)
ESM 1 (DOCX 67 kb)
10811_2018_1716_MOESM2_ESM.pdf (273 kb)
ESM 2 (PDF 273 kb)


  1. Abdel-Raouf N, Al-Homaidan AA, Ibraheem IBM (2012) Microalgae and wastewater treatment. Saudi J Biol Sci 19:257–275CrossRefGoogle Scholar
  2. Ahemad M (2014) Bacterial mechanisms for Cr(VI) resistance and reduction: an overview and recent advances. Folia Microbiol 59:321–332CrossRefGoogle Scholar
  3. APHA (2005) Standard methods for the examination of water and wastewater. 21st edition. By Clesceri LS, Greenberg AE, Eaton AD, American Public Health Association, American Water Works Association and Water Pollution Control Federation, Washington DCGoogle Scholar
  4. Arita A, Costa M (2011) Environmental agents and epigenetics. In: Tollefsbol T (ed) Handbook of epigenetics. Academic Press, NY, pp 459–476CrossRefGoogle Scholar
  5. Arun N, Singh DP (2014) Chromium (VI) induced oxidative stress in halotolerant alga Dunaliella salina and D. tertiolecta isolated from Ssambhar salt lake of Rajasthan (India). Cell Mol Biol 60:90–96PubMedGoogle Scholar
  6. Barrera-Díaz CE, Lugo-Lugo V, Bilyeu B (2012) A review of chemical, electrochemical and biological methods for aqueous Cr(VI) reduction. J Hazard Mater 223:1–12CrossRefGoogle Scholar
  7. Bell WH, Sakshaug E (1980) Bacterial utilization of algal extracellular products. 2. A kinetic study of natural populations. Limnol Oceanogr 25:1021–1033CrossRefGoogle Scholar
  8. Bridgewater LC, Manning FC, Patierno SR (1994) Base-specific arrest of in vitro DNA replication by carcinogenic chromium: relationship to DNA interstrand crosslinking. Carcinogenesis 15:2421–2427CrossRefGoogle Scholar
  9. Bruckner CG, Bahulikar R, Rahalkar M, Schink B, Kroth PG (2008) Bacteria associated with benthic diatoms from Lake Constance: phylogeny and influences on diatom growth and secretion of extracellular polymeric substances. Appl Environ Microbiol 74:7740–7749CrossRefGoogle Scholar
  10. Bush MB (2003) Ecology of a Changing Planet, 3rd Edition. Prentice Hall, New JerseyGoogle Scholar
  11. CCAP (2015) The Culture Collection of Algae and Protozoa. Medium recipe for 3N-BBM+V (Bold Basal Medium with 3-fold Nitrogen and Vitamins; modified). Accessed 27 Mar 2017
  12. Chen JM, Hao OJ (1998) Microbial chromium (VI) reduction. Crit Rev Environ Sci Technol 28:219–251CrossRefGoogle Scholar
  13. Chirwa EM, Molokwane PE (2011) Biological Cr (VI) reduction: microbial diversity, kinetics and biotechnological solutions to pollution. In: Solo A (ed) Biodiversity. InTech, Riejeka, pp 75–100Google Scholar
  14. Cicci A, Sed G, Bravi M (2017) Potential of choline chloride-based natural deep eutectic solvents (nades) in the extraction of microalgal metabolites. Chem Eng Trans 57:61–66Google Scholar
  15. Coenye T, Falsen E, Vancanneyt M, Hoste B, Govan JRW, Kersters K, Vandamme P (1999) Classification of Alcaligenes faecalis-like isolates from the environment and human clinical samples as Ralstonia gilardii sp. nov. Int J Syst Bacteriol 49:405–413CrossRefGoogle Scholar
  16. Cole JJ, Likens GE, Strayer DL (1982) Photosynthetically produced dissolved organic carbon: an important carbon source for planktonic bacteria. Limnol Oceanogr 27:1080–1090CrossRefGoogle Scholar
  17. Dakhama A, De la Noüe J, Lavoie MC (1993) Isolation and identification of antialgal substances produced by Pseudomonas aeruginosa. J Appl Phycol 5:297–306CrossRefGoogle Scholar
  18. Dong T, Knoshaug EP, Pienkos PT, Laurens LM (2016) Lipid recovery from wet oleaginous microbial biomass for biofuel production: a critical review. Appl Energy 177:879–895CrossRefGoogle Scholar
  19. EPA, US (1998) Toxicological review of hexavalent chromium. Washington, DCGoogle Scholar
  20. EPA-Odessa (2005) Pump and treat and in situ chemical treatment of contaminated groundwater at the Odessa Chromium II South Plume Superfund Site Odessa, Ector County, Texas. U.S. Environmental Protection AgencyGoogle Scholar
  21. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  22. Fujie K, Toda K, Ohtake H (1990) Bacterial reduction of toxic hexavalent chromium using a fed-batch culture of Enterobacter cloacae strain HO1. J Ferment Bioengr 69:365–367CrossRefGoogle Scholar
  23. Gardiner M, Hoke DE, Egan S (2014) An ortholog of the Leptospira interrogans lipoprotein LipL32 aids in the colonization of Pseudoalteromonas tunicata to host surfaces. Front Microbiol 5:323CrossRefGoogle Scholar
  24. Glauert AM (1975) Fixation, dehydration and embedding of biological specimens. No. 04; QH327, G5Google Scholar
  25. Grima EM, Belarbi EH, Fernández FA, Medina AR, Chisti Y (2003) Recovery of microalgal biomass and metabolites: process options and economics. Biotechnol Adv 20:491–515CrossRefGoogle Scholar
  26. Guo Z, Tong YW (2014) The interactions between Chlorella vulgaris and algal symbiotic bacteria under photoautotrophic and photoheterotrophic conditions. J Appl Phycol 26:1483–1492CrossRefGoogle Scholar
  27. Gutiérrez-Corona JF, Romo-Rodríguez P, Santos-Escobar F, Espino-Saldana AE, Hernández-Escoto H (2016) Microbial interactions with chromium: basic biological processes and applications in environmental biotechnology. World J Microbiol Biotechnol 32:191CrossRefGoogle Scholar
  28. Hassoun EA, Stohs SJ (1995) Chromium-induced production of reactive oxygen species, DNA single-strand breaks, nitric oxide production, and lactate dehydrogenase leakage in J774A. 1 cell cultures. J Biochem Mol Toxicol 10:315–321CrossRefGoogle Scholar
  29. Igboamalu T, Chirwa E (2016) Kinetic study of Cr(VI) reduction in an indigenous mixed culture of bacteria in the presence of as(III). Chem Eng Trans 49:439–444Google Scholar
  30. Ji RP, Lu XW, Li XN, Pu YP (2009) Biological degradation of algae and microcystins by microbial enrichment on artificial media. Ecol Eng 35:1584–1588CrossRefGoogle Scholar
  31. Kalckar HM (1974) Origins of the concept oxidative phosphorylation. Mol Cell Biochem 5:55–63CrossRefGoogle Scholar
  32. Kang SY, Lee JU, Kim KW (2007) Biosorption of Cr(III) and Cr(VI) onto the cell surface of Pseudomonas aeruginosa. Biochem Eng J 36:54–58CrossRefGoogle Scholar
  33. Kleinová A, Cvengrošová Z, Rimarčík J, Buzetzki E, Mikulec J, Cvengroš J (2012) Biofuels from algae. Procedia Eng 42:231–238CrossRefGoogle Scholar
  34. Kothari R, Pathak VV, Kumar V, Singh DP (2012) Experimental study for growth potential of unicellular alga Chlorella pyrenoidosa on dairy waste water: an integrated approach for treatment and biofuel production. Bioresour Technol 116:466–470CrossRefGoogle Scholar
  35. Kratochvil D, Pimentel P, Volesky B (1998) Removal of trivalent and hexavalent chromium by seaweed biosorbent. Environ Sci Technol 32:2693–2698CrossRefGoogle Scholar
  36. Liang Z, Liu Y, Ge F, Xu Y, Tao N, Peng F, Wong M (2013) Efficiency assessment and pH effect in removing nitrogen and phosphorus by algae-bacteria combined system of Chlorella vulgaris and Bacillus licheniformis. Chemosphere 92:1383–1389CrossRefGoogle Scholar
  37. Lloyd JR (2003) Microbial reduction of metals and radionuclides. FEMS Microbiol Rev 27:411–425CrossRefGoogle Scholar
  38. Loukidou MX, Zouboulis AI, Karapantsios TD, Matis KA (2004) Equilibrium and kinetic modeling of chromium (VI) biosorption by Aeromonas caviae. Colloids Surf A 242:93–104CrossRefGoogle Scholar
  39. Macfie A, Hagan E, Zhitkovich A (2009) Mechanism of DNA− protein cross-linking by chromium. Chem Res Toxicol 23:341–347CrossRefGoogle Scholar
  40. Marande W, López-García P, Moreira D (2009) Eukaryotic diversity and phylogeny using small-and large-subunit ribosomal RNA genes from environmental samples. Environ Microbiol 11:3179–3188CrossRefGoogle Scholar
  41. Miranda J, Krishnakumar G, Gonsalves R (2012) Cr6+ bioremediation efficiency of Oscillatoria laete-virens (Crouan & Crouan) Gomont and Oscillatoria trichoides Szafer: kinetics and equilibrium study. J Appl Phycol 24:1439–1454CrossRefGoogle Scholar
  42. Molokwane PE, Nkhalambayausi-Chirwa EM, Meli KC (2008) Chromium (VI) reduction in activated sludge bacteria exposed to high chromium loading: Brits culture (South Africa). Water Res 42:4538–4548CrossRefGoogle Scholar
  43. Munoz R, Guieysse B (2006) Algal–bacterial processes for the treatment of hazardous contaminants: a review. Water Res 40:2799–2815CrossRefGoogle Scholar
  44. Murphy V, Hughes H, McLoughlin P (2008) Comparative study of chromium biosorption by red, green and brown seaweed biomass. Chemosphere 70:1128–1134CrossRefGoogle Scholar
  45. Nakayama T, Watanabe S, Mitsui K, Uchida H, Inouye I (1996) The phylogenetic relationship between the Chlamydomonadales and Chlorococcales inferred from 18S rDNA sequence data. Phycol Res 44:47–55CrossRefGoogle Scholar
  46. Nelson YM, Lion LW, Shuler ML, Ghiorse WC (1996) Modeling oligotrophic biofilm formation and lead adsorption to biofilm components. Environ Sci Technol 30:2027–2035CrossRefGoogle Scholar
  47. Ozdemir G, Ceyhan N, Ozturk T, Akirmak F, Cosar T (2004) Biosorption of chromium (VI), cadmium (II) and copper (II) by Pantoea sp. TEM18 Chem Eng J 102:249–253CrossRefGoogle Scholar
  48. Pell L, Löhn S, Weinberger G, Kuchta K, Hanelt D (2017) Mild disintegration methods of microalgae–bacteria flocs from wastewater treatment. J Appl Phycol 29:843–851CrossRefGoogle Scholar
  49. Pritchard DE, Singh J, Carlisle DL, Patierno SR (2000) Cyclosporin a inhibits chromium(VI)-induced apoptosis and mitochondrial cytochrome c release and restores clonogenic survival in CHO cells. Carcinogenesis 21:2027–2033CrossRefGoogle Scholar
  50. Ramanan R, Kim BH, Cho DH, Oh HM, Kim HS (2016) Algae–bacteria interactions: evolution, ecology and emerging applications. Biotechnol Adv 34:14–29CrossRefGoogle Scholar
  51. Rodríguez MC, Barsanti L, Passarelli V, Evangelista V, Conforti V, Gualtieri P (2007) Effects of chromium on photosynthetic and photoreceptive apparatus of the alga Chlamydomonas reinhardtii. Environ Res 105:234–239CrossRefGoogle Scholar
  52. Şahin Y, Öztürk A (2005) Biosorption of chromium (VI) ions from aqueous solution by the bacterium Bacillus thuringiensis. Process Biochem 40:1895–1901CrossRefGoogle Scholar
  53. Shen H, Wang YT (1994) Modeling hexavalent chromium reduction in Escherichia coli ATCC 33456. Biotechnol Bioeng 43:293–300CrossRefGoogle Scholar
  54. Sibi G (2016) Biosorption of chromium from electroplating and galvanizing industrial effluents under extreme conditions using Chlorella vulgaris. Green Energy Environ 1:172–177CrossRefGoogle Scholar
  55. Singh J, Bridgewater LC, Patierno SR (1998a) Differential sensitivity of chromium-mediated DNA interstrand crosslinks and DNA–protein crosslinks to disruption by alkali and EDTA. Toxicol Sci 45:72–76PubMedGoogle Scholar
  56. Singh J, McLean JA, Pritchard DE, Montaser A, Patierno SR (1998b) Sensitive quantitation of chromium-DNA adducts by inductively coupled plasma mass spectrometry with a direct injection high-efficiency nebulizer. Toxicol Sci 46:260–265PubMedGoogle Scholar
  57. Smith WA, Apel WA, Petersen JN, Peyton BM (2002) Effect of carbon and energy source on bacterial chromate reduction. Bioremediat J 6:205–215CrossRefGoogle Scholar
  58. Szymczak-Żyla M, Kowalewska G, Louda JW (2008) The influence of microorganisms on chlorophyll a degradation in the marine environment. Limnol Oceanogr 53:851–862CrossRefGoogle Scholar
  59. Thatoi H, Das S, Mishra J, Rath BP, Das N (2014) Bacterial chromate reductase, a potential enzyme for bioremediation of hexavalent chromium: a review. J Environ Manag 146:383–399CrossRefGoogle Scholar
  60. Thomas PG, Quinn PJ, Williams WP (1985) The origin of photosystem-I mediated electron transport stimulation in heat-stressed chloroplasts. Planta 167:133–139CrossRefGoogle Scholar
  61. Toncheva-Panova T, Ivanova J (2000) Influence of physiological factors on the lysis effect of Cytophaga on the red microalga Rhodella reticulata. J Appl Microbiol 88:358–363CrossRefGoogle Scholar
  62. Tsou TC, Chen CL, Liu TY, Yang JL (1996) Induction of 8-hydroxydeoxyguanosine in DNA by chromium(III) plus hydrogen peroxide and its prevention by scavengers. Carcinogenesis 17:103–108CrossRefGoogle Scholar
  63. Ünal D, Işik NO, Sukatar A (2010) Effects of Chromium VI stress on green alga Ulva lactuca (L.). Turk J Biol 34:119–124Google Scholar
  64. Vidotti A, Coelho R, Franco LM, Franco TT (2014) Miniaturized culture for heterotrophic microalgae using low cost carbon sources as a tool to isolate fast and economical strains. Chem Eng Trans 38:325–330Google Scholar
  65. Viti C, Marchi E, Decorosi F, Giovannetti L (2014) Molecular mechanisms of Cr(VI) resistance in bacteria and fungi. FEMS Microbiol Rev 38:633–659CrossRefGoogle Scholar
  66. Volland S, Lütz C, Michalke B, Lütz-Meindl U (2012) Intracellular chromium localization and cell physiological response in the unicellular alga Micrasterias. Aquat Toxicol 109:59–69CrossRefGoogle Scholar
  67. Wise JP, Leonard JC, Patierno SR (1992) Clastogenicity of lead chromate particles in hamster and human cells. Mutat Res 278:69–79CrossRefGoogle Scholar
  68. World Health Organization (2004) Guidelines for drinking-water quality: recommendations (Vol. 1). World Health Organization, GenevaGoogle Scholar
  69. Xu J, Bubley GJ, Detrick B, Blankenship LJ, Patierno SR (1996) Chromium(VI) treatment of normal human lung cells results in guanine-specific DNA polymerase arrest, DNA–DNA cross-links and S-phase blockade of cell cycle. Carcinogenesis 17:1511–1517CrossRefGoogle Scholar
  70. Xue Y, Jin W, Du H, Zheng S, Sun Z, Yan W, Zhang Y (2016) Electrochemical Cr (III) oxidation and mobilization by in situ generated reactive oxygen species in alkaline solution. J Electrochem Soc 163:684–689CrossRefGoogle Scholar
  71. Yewalkar SN, Dhumal KN, Sainis JK (2007) Chromium (VI)-reducing Chlorella spp. isolated from disposal sites of paper-pulp and electroplating industry. J Appl Phycol 19:459–465CrossRefGoogle Scholar
  72. Zhiguo H, Fengling G, Tao S, Yuehua H, Chao H (2009) Isolation and characterization of a Cr(VI)-reduction Ochrobactrum sp. strain CSCr-3 from chromium landfill. J Hazard Mater 163:869–873CrossRefGoogle Scholar
  73. Ziagova M, Dimitriadis G, Aslanidou D, Papaioannou X, Tzannetaki EL, Liakopoulou-Kyriakides M (2007) Comparative study of CD (II) and Cr (VI) biosorption on Staphylococcus xylosus and Pseudomonas sp. in single and binary mixtures. Bioresour Technol 98:2859–2865CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Chemical EngineeringUniversity of PretoriaPretoriaSouth Africa

Personalised recommendations