Skip to main content
SpringerLink
Log in

Application of Cell Immobilization in Slurry-Phase Bioremediation: Phenanthrene Biodegradation and Detoxification

  • Protocol
  • First Online:
Toxicity and Biodegradation Testing

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

Abstract

Polycyclic aromatic hydrocarbons (PAHs) are among the detrimental soil contaminants worldwide. Free microbial cells have been widely used for PAHs biodegradation in contaminated soils. However, only few studies have been carried out on the application of the immobilized cell system for bioremediation of PAH contaminated soil/slurry. In this chapter, we reviewed the literature on PAHs biodegradation, especially in solid/slurry phase by immobilized cell systems. This was followed by our experimental study on bioremediation of a clayey soil contaminated with phenantherene, a model contaminant, at 250, 500, 1000, and 2000 (mg PHE)(kg dry soil)−1 using immobilized cell (IC) and free cell (FC). Formation of intermediate metabolites (IMs) of the PAH oxidation pathways was also examined since some of the IMs could be reluctant to further biodegradation or be more toxic than the parent pollutants. This was performed by measuring the total organic carbon (TOC) of soil and chemical oxygen demand (COD) of aqueous phase. Additionally, the effect of temperature (10, 20, 30, 40 °C), pH (5, 7, 9), and inoculum size (300 and 600 mg/L) on IC and FC systems was investigated. According to t-test results, immobilization had an insignificant effect on PHE biodegradation up to 1000 mg PHE (kg dry soil)−1. However, less accumulation of IMs was observed at higher PHE content of soil for IC as compared to FC system, showing the superiority of immobilization. TOC analysis also showed the oxidation of PHE in soil samples into nontoxic intermediate metabolites that in turn were mineralized by our microbial consortium. Maximum PHE biodegradation was obtained at 30 °C, pH of 7, initial 2000 (mg PHE)(kg dry soil)−1 and 600 mg L−1 of inoculum for both the systems. However, it was shown that the IC system was tolerable to acidic condition with retaining 85% of its biodegradation capability at neutral pH while the FC system was highly affected by acidic pH and retained only 33% of its capability. Our results show the superiority of cell immobilization compared to free cell system, especially at high concentration of PHE and under harsh environmental conditions. This can have generic application for other PAHs.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Bamforth SM, Singleton I (2005) Bioremediation of polycyclic aromatic hydrocarbons: current knowledge and future directions. J Chem Technol Biotechnol 80(7):723–736

    Article  CAS  Google Scholar 

  2. Boonchan S, Britz ML, Stanley GA (2000) Degradation and mineralization of high-molecular-weight polycyclic aromatic hydrocarbons by defined fungal-bacterial cocultures. Appl Environ Microbiol 66(3):1007–1019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Lu XY, Zhang T, Fang HHP (2011) Bacteria-mediated PAH degradation in soil and sediment. Appl Microb Biotechnol 89(5):1357–1371

    Article  CAS  Google Scholar 

  4. Haritash AK, Kaushik CP (2009) Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): a review. J Hazard Mater 169(1–3):1–15

    Article  CAS  PubMed  Google Scholar 

  5. Juhasz AL, Naidu R (2000) Bioremediation of high molecular weight polycyclic aromatic hydrocarbons: a review of the microbial degradation of benzo[a]pyrene. Int Biodeter Biodegrad 45(1–2):57–88

    Article  CAS  Google Scholar 

  6. Singh A, Kuhad RC, Ward OP (2009) Advances in applied bioremediation

    Google Scholar 

  7. Martienssen M, Grundl M (2000) Immobilized microorganisms for the degradation of PAH and hydrocarbons from contaminated soil. Chem Eng Technol 23(10):860–863

    Article  CAS  Google Scholar 

  8. Partovinia A, Naeimpoor F, Hejazi P (2010) Carbon content reduction in a model reluctant clayey soil: slurry phase n-hexadecane bioremediation. J Hazard Mater 181:133–139

    Article  CAS  PubMed  Google Scholar 

  9. Weissenfels W, Klewer H-J, Langhoff J (1992) Adsorption of polycyclic aromatic hydrocarbons (PAHs) by soil particles: influence on biodegradability and biotoxicity. Appl Microbiol Biotechnol 36(5):689–696

    Article  CAS  PubMed  Google Scholar 

  10. Yuan SY, Shiung LC, Chang BV (2002) Biodegradation of polycyclic aromatic hydrocarbons by inoculated microorganisms in soil. Bull Environ Contam Toxicol 69(1):66–73

    Article  CAS  PubMed  Google Scholar 

  11. Masy T, Demanèche S, Tromme O, Thonart P, Jacques P, Hiligsmann S, Vogel TM (2016) Hydrocarbon biostimulation and bioaugmentation in organic carbon and clay-rich soils. Soil Biol Biochem 99:66–74

    Article  CAS  Google Scholar 

  12. Okuda T, Alcántara-Garduño ME, Suzuki M, Matsui C, Kose T, Nishijima W, Okada M (2007) Enhancement of biodegradation of oil adsorbed on fine soils in a bioslurry reactor. Chemosphere 68(2):281–286

    Article  CAS  PubMed  Google Scholar 

  13. Zheng Z, Obbard JP (2001) Effect of non-ionic surfactants on elimination of polycyclic aromatic hydrocarbons (PAHs) in soil-slurry by Phanerochaete chrysosporium. J Chem Technol Biotechnol 76(4):423–429

    Article  CAS  Google Scholar 

  14. Colombo M, Cavalca L, Bernasconi S, Andreoni V (2011) Bioremediation of polyaromatic hydrocarbon contaminated soils by native microflora and bioaugmentation with Sphingobium chlorophenolicum strain C3R: a feasibility study in solid- and slurry-phase microcosms. Int Biodeter Biodegrad 65(1):191–197

    Article  CAS  Google Scholar 

  15. Woo S, Jeon C, Park J (2004) Phenanthrene biodegradation in soil slurry systems: influence of salicylate and triton X-100. Korean J Chem Eng 21(2):412–418

    Article  CAS  Google Scholar 

  16. Di Gennaro P, Franzetti A, Bestetti G, Lasagni M, Pitea D, Collina E (2008) Slurry phase bioremediation of PAHs in industrial landfill samples at laboratory scale. Waste Manag 28(8):1338–1345

    Article  PubMed  Google Scholar 

  17. Launen LA, Buggs VH, Eastep ME, Enriquez RC, Leonard JW, Blaylock MJ, Huang J-W, Häggblom MM (2002) Bioremediation of Polyaromatic hydrocarbon-contaminated sediments in aerated bioslurry reactors. Biorem J 6(2):125–141

    Article  CAS  Google Scholar 

  18. Doick KJ, Semple KT (2003) The effect of soil:water ratios on the mineralisation of phenanthrene:LNAPL mixtures in soil. FEMS Microbiol Lett 220(1):29–33

    Article  CAS  PubMed  Google Scholar 

  19. Brown DG, Guha S, Jaffé PR (1999) Surfactant-enhanced biodegradation of a PAH in soil slurry reactors. Biorem J 3(3):269–283

    Article  CAS  Google Scholar 

  20. Kim IS, Park JS, Kim KW (2001) Enhanced biodegradation of polycyclic aromatic hydrocarbons using nonionic surfactants in soil slurry. Appl Geochem 16(11–12):1419–1428

    Article  CAS  Google Scholar 

  21. Gottfried A, Singhal N, Elliot R, Swift S (2010) The role of salicylate and biosurfactant in inducing phenanthrene degradation in batch soil slurries. Appl Microbiol Biotechnol 86(5):1563–1571

    Article  CAS  PubMed  Google Scholar 

  22. Li PJ, Wang X, Stagnitti F, Li L, Gong ZQ, Zhang HR, Xiong XZ, Austin C (2005) Degradation of phenanthrene and pyrene in soil slurry reactors with immobilized bacteria Zoogloea sp. Environ Eng Sci 22(3):390–399

    Article  CAS  Google Scholar 

  23. Venkata Mohan S, Ramakrishna M, Shailaja S, Sarma PN (2007) Influence of soil-water ratio on the performance of slurry phase bioreactor treating herbicide contaminated soil. Bioresour Technol 98(13):2584–2589

    Article  CAS  PubMed  Google Scholar 

  24. Boopathy R (2000) Factors limiting bioremediation technologies. Bioresour Technol 74(1):63–67

    Article  CAS  Google Scholar 

  25. Zhang GY, Ling JY, Sun HB, Luo J, Fan YY, Cui ZJ (2009) Isolation and characterization of a newly isolated polycyclic aromatic hydrocarbons-degrading Janibacter anophelis strain JY11. J Hazard Mater 172(2–3):580–586

    Article  CAS  PubMed  Google Scholar 

  26. Kim J-D, Shim S-H, Lee C-G (2005) Degradation of phenanthrene by bacterial strains isolated from soil in oil refinery fields in Korea. J Microbiol Biotechnol 15(2):337–345

    CAS  Google Scholar 

  27. Zhao H-P, Wu Q-S, Wang L, Zhao X-T, Gao H-W (2009) Degradation of phenanthrene by bacterial strain isolated from soil in oil refinery fields in Shanghai China. J Hazard Mater 164(2–3):863–869

    Article  CAS  PubMed  Google Scholar 

  28. Chang BV, Shiung LC, Yuan SY (2002) Anaerobic biodegradation of polycyclic aromatic hydrocarbon in soil. Chemosphere 48(7):717–724

    Article  CAS  PubMed  Google Scholar 

  29. Kastner M, Breuer-Jammali M, Mahro B (1998) Impact of inoculation protocols, salinity, and pH on the degradation of polycyclic aromatic hydrocarbons (PAHs) and survival of PAH-degrading bacteria introduced into soil. Appl Environ Microbiol 64(1):359–362

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Kim H, Lindsay K, Pfaender F (2008) Enhanced mobilization of field contaminated soil-bound PAHs to the aqueous phase under anaerobic conditions. Water Air Soil Pollut 189(1):135–147

    Article  CAS  Google Scholar 

  31. Chauhan A, Fazlurrahman OJ, Jain R (2008) Bacterial metabolism of polycyclic aromatic hydrocarbons: strategies for bioremediation. Indian J Microbiol 48(1):95–113

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kim Y-H, Freeman JP, Moody JD, Engesser K-H, Cerniglia CE (2005) Effects of pH on the degradation of phenanthrene and pyrene by Mycobacterium vanbaalenii PYR-1. Appl Microbiol Biotechnol 67(2):275–285

    Article  CAS  PubMed  Google Scholar 

  33. Mrozik A, Piotrowska-Seget Z (2010) Bioaugmentation as a strategy for cleaning up of soils contaminated with aromatic compounds. Microbiol Res 165(5):363–375

    Article  CAS  PubMed  Google Scholar 

  34. Kim HS, Weber WJ (2005) Polycyclic aromatic hydrocarbon behavior in bioactive soil slurry reactors amended with a nonionic surfactant. Environ Toxicol Chem 24(2):268–276

    Article  CAS  PubMed  Google Scholar 

  35. Cassidy MB, Mullineers H, Lee H, Trevors JT (1997) Mineralization of pentachlorophenol in a contaminated soil by Pseudomonas sp UG30 cells encapsulated in κ-carrageenan. J Ind Microbiol Biotechnol 19(1):43–48

    Article  CAS  Google Scholar 

  36. Chen B, Yuan M, Qian L (2012) Enhanced bioremediation of PAH-contaminated soil by immobilized bacteria with plant residue and biochar as carriers. J Soils Sediments 12(9):1350–1359

    Article  CAS  Google Scholar 

  37. Wang S, Li X, Liu W, Li P, Kong L, Ren W, Wu H, Tu Y (2012) Degradation of pyrene by immobilized microorganisms in saline-alkaline soil. J Environ Sci 24(9):1662–1669

    Article  CAS  Google Scholar 

  38. Weir S, Dupuis S, Providenti M, Lee H, Trevors J (1995) Nutrient-enhanced survival of and phenanthrene mineralization by alginate-encapsulated and free Pseudomonas sp. UG14Lr cells in creosote-contaminated soil slurries. Appl Microbiol Biotechnol 43(5):946–951

    Article  CAS  PubMed  Google Scholar 

  39. Weir SC, Providenti MA, Lee H, Trevors JT (1996) Effect of alginate encapsulation and selected disinfectants on survival of and phenanthrene mineralization by Pseudomonas sp UG14Lr in creosote-contaminated soil. J Ind Microbiol 16(1):62–67

    Article  CAS  Google Scholar 

  40. Wang X, Gong ZQ, Li PJ, Zhang LH (2007) Degradation of pyrene in soils by free and immobilized yeasts, Candida tropicals. Bull Environ Contam Toxicol 78(6):522–526

    Article  CAS  PubMed  Google Scholar 

  41. Somtrakoon K, Suanjit S, Pokethitiyook P, Kruatrachue M, Cassidy MB, Trevors JT, Lee H, Upatham S (2009) Comparing phenanthrene degradation by alginate-encapsulated and free Pseudomonas sp UG14Lr cells in heavy metal contaminated soils. J Chem Technol Biotechnol 84(11):1660–1668

    Article  CAS  Google Scholar 

  42. R-y H, W-j T, Liu Q, Yu H-b, Jin X, Y-g Z, Y-h Z, Feng G (2016) Enhanced biodegradation of pyrene and indeno(1,2,3-cd)pyrene using bacteria immobilized in cinder beads in estuarine wetlands. Mar Poll Bull 102(1):128–133

    Article  Google Scholar 

  43. Guerin W, Jones G (1988) Two-stage mineralization of phenanthrene by estuarine enrichment cultures. Appl Environ Microbiol 54(4):929–936

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Silva IS, dos Santos ED, de Menezes CR, de Faria AF, Franciscon E, Grossman M, Durrant LR (2009) Bioremediation of a polyaromatic hydrocarbon contaminated soil by native soil microbiota and bioaugmentation with isolated microbial consortia. Bioresour Technol 100(20):4669–4675

    Article  CAS  PubMed  Google Scholar 

  45. Xu Y, Lu M (2010) Bioremediation of crude oil-contaminated soil: comparison of different biostimulation and bioaugmentation treatments. J Hazard Mater 183(1–3):395–401. doi:10.1016/j.jhazmat.2010.07.038

    Article  CAS  PubMed  Google Scholar 

  46. Park KS, Sims RC, Dupont RR, Doucette WJ, Matthews JE (1990) Fate of PAH compounds in two soil types: influence of volatilization, abiotic loss and biological activity. Environ Toxicol Chem 9(2):187–195. doi:10.1002/etc.5620090208

    Article  CAS  Google Scholar 

  47. Lozinsky VI, Zubov AL, Titova EF (1997) Poly(vinyl alcohol) cryogels employed as matrices for cell immobilization. 2. Entrapped cells resemble porous fillers in their effects on the properties of PVA-cryogel carrier. Enzym Microb Technol 20(3):182–190

    Article  CAS  Google Scholar 

  48. Woo SH, Park JM (1999) Evaluation of drum bioreactor performance used for decontamination of soil polluted with polycyclic aromatic hydrocarbons. J Chem Technol Biotechnol 74(10):937–944

    Article  CAS  Google Scholar 

  49. Box JD (1983) Investigation of the Folin-Ciocalteau phenol reagent for the determination of polyphenolic substances in natural waters. Water Res 17(5):511–525

    Article  CAS  Google Scholar 

  50. Samanta SK, Chakraborti AK, Jain RK (1999) Degradation of phenanthrene by different bacteria: evidence for novel transformation sequences involving the formation of 1-naphthol. Appl Microbiol Biotechnol 53(1):98–107

    Article  CAS  PubMed  Google Scholar 

  51. Schulte EE (1995) Recommended soil organic matter tests. In: Horton ML (ed) Recommended soil testing procedures for the Northeastern United States,vol 493, 2nd edn. Northeastern Regional Publication, Columbia, MO

    Google Scholar 

  52. Clesceri LS, Greenberg AE, Eaton AD (1998) Standard methods for the examination of water and wastewater, 20th edn. APHA American Public Health Association, Washington, DC

    Google Scholar 

  53. Mulla SI, Talwar MP, Bagewadi ZK, Hoskeri RS, Ninnekar HZ (2013) Enhanced degradation of 2-nitrotoluene by immobilized cells of micrococcus sp. strain SMN-1. Chemosphere 90(6):1920–1924

    Article  CAS  PubMed  Google Scholar 

  54. Lin M, Liu Y, Chen W, Wang H, Hu X (2014) Use of bacteria-immobilized cotton fibers to absorb and degrade crude oil. Int Biodeter Biodegrad 88(0):8–12

    Article  CAS  Google Scholar 

  55. Feijoo-Siota L, Rosa-Dos-Santos F, de Miguel T, Villa TG (2008) Biodegradation of naphthalene by Pseudomonas stutzeri in marine environments: testing cells entrapment in calcium alginate for use in water detoxification. Biorem J 12(4):185–192

    Article  CAS  Google Scholar 

  56. Zinjarde SS, Pant A (2000) Crude oil degradation by free and immobilized cells of Yarrowia lipolytica NCIM 3589. J Environ Sci Health A Tox Hazard Subst Environ Eng 35(5):755–763

    Article  Google Scholar 

  57. Jianlong W, Liping H, Hanchang S, Yi Q (2001) Biodegradation of quinoline by gel immobilized Burkholderia sp. Chemosphere 44(5):1041–1046

    Article  CAS  PubMed  Google Scholar 

  58. Manohar S, Kim CK, Karegoudar TB (2001) Enhanced degradation of naphthalene by immobilization of Pseudomonas sp strain NGK1 in polyurethane foam. Appl Microbiol Biotechnol 55(3):311–316

    Article  CAS  PubMed  Google Scholar 

  59. Moslemy P, Neufeld RJ, Guiot SR (2002) Biodegradation of gasoline by gellan gum-encapsulated bacterial cells. Biotechnol Bioeng 80(2):175–184

    Article  CAS  PubMed  Google Scholar 

  60. Partovinia A, Naeimpoor F (2013) Phenanthrene biodegradation by immobilized microbial consortium in polyvinyl alcohol cryogel beads. Int Biodeter Biodegrad 85:337–344

    Article  CAS  Google Scholar 

  61. Stringfellow WT, Alvarez-Cohen L (1999) Evaluating the relationship between the sorption of PAHs to bacterial biomass and biodegradation. Water Res 33(11):2535–2544

    Article  CAS  Google Scholar 

  62. Nano G, Borroni A, Rota R (2003) Combined slurry and solid-phase bioremediation of diesel contaminated soils. J Hazard Mater 100(1–3):79–94

    Article  CAS  PubMed  Google Scholar 

  63. Noordman WH, Wachter JHJ, de Boer GJ, Janssen DB (2002) The enhancement by surfactants of hexadecane degradation by Pseudomonas aeruginosa varies with substrate availability. J Biotechnol 94(2):195–212

    Article  CAS  PubMed  Google Scholar 

  64. Leys N, Bastiaens L, Verstraete W, Springael D (2005) Influence of the carbon/nitrogen/phosphorus ratio on polycyclic aromatic hydrocarbon degradation by Mycobacterium and Sphingomonas in soil. Appl Microbiol Biotechnol 66(6):726–736

    Article  CAS  PubMed  Google Scholar 

  65. Su D, Li PJ, Stagnitti F, Xiong XZ (2006) Biodegradation of benzo[a]pyrene in soil by Mucor sp SF06 and Bacillus sp SB02 co-immobilized on vermiculite. J Environ Sci 18(6):1204–1209

    Article  CAS  Google Scholar 

  66. Brinda Lakshmi M, Muthukumar K, Velan M (2012) Immobilization of Mycoplana sp. MVMB2 isolated from petroleum contaminated soil onto papaya stem (Carica Papaya L.) and its application on degradation of Phenanthrene. Clean (Weinh) 40(8):870–877

    CAS  Google Scholar 

  67. Masakorala K, Yao J, Cai M, Chandankere R, Yuan H, Chen H (2013) Isolation and characterization of a novel phenanthrene (PHE) degrading strain Psuedomonas sp. USTB-RU from petroleum contaminated soil. J Hazard Mater 263 pt 2:493–500

    Article  PubMed  Google Scholar 

  68. Cassidy MB, Lee H, Trevors JT (1996) Environmental applications of immobilized microbial cells: a review. J Ind Microbiol 16(2):79–101

    Article  CAS  Google Scholar 

  69. Wang M-Y, Yu Y-T, Chang TMS (2005) New method for preparing more stable microcapsules for the entrapment of genetically engineered cells. Artif Cell Blood Sub 33(3):257–269

    Article  CAS  Google Scholar 

  70. Eweis JB, Eragas SJ, Chang DPY, Schroeder ED (1998) Bioremediation principles. McGraw-Hill Companies, Boston, MA

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biotechnology Research Laboratory, School of Chemical Engineering, Iran University of Science and Technology, Tehran, Iran

    Ali Partovinia & Fereshteh Naeimpoor

Authors
  1. Ali Partovinia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fereshteh Naeimpoor
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fereshteh Naeimpoor .

Editor information

Editors and Affiliations

  1. Biosciences Institute, São Paulo State University (UNESP), Rio Claro, São Paulo, Brazil

    Ederio Dino Bidoia

  2. Biosciences Institute, São Paulo State University (UNESP), Rio Claro, São Paulo, Brazil

    Renato Nallin Montagnolli

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Partovinia, A., Naeimpoor, F. (2018). Application of Cell Immobilization in Slurry-Phase Bioremediation: Phenanthrene Biodegradation and Detoxification. In: Bidoia, E., Montagnolli, R. (eds) Toxicity and Biodegradation Testing. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7425-2_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7425-2_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7424-5

  • Online ISBN: 978-1-4939-7425-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation