Applied Microbiology and Biotechnology

, Volume 96, Issue 1, pp 231–240 | Cite as

Silica gel-encapsulated AtzA biocatalyst for atrazine biodegradation

  • Eduardo Reátegui
  • Erik Reynolds
  • Lisa Kasinkas
  • Amit Aggarwal
  • Michael J. Sadowsky
  • Alptekin Aksan
  • Lawrence P. Wackett
Environmental biotechnology

Abstract

Encapsulation of recombinant Escherichia coli cells expressing a biocatalyst has the potential to produce stable, long-lasting enzyme activity that can be used for numerous applications. The current study describes the use of this technology with recombinant E. coli cells expressing the atrazine-dechlorinating enzyme AtzA in a silica/polymer porous gel. This novel recombinant enzyme-based method utilizes both adsorption and degradation to remove atrazine from water. A combination of silica nanoparticles (Ludox TM40), alkoxides, and an organic polymer was used to synthesize a porous gel. Gel curing temperatures of 23 or 45 °C were used either to maintain cell viability or to render the cells non-viable, respectively. The enzymatic activity of the encapsulated viable and non-viable cells was high and extremely stable over the time period analyzed. At room temperature, the encapsulated non-viable cells maintained a specific activity between (0.44 ± 0.06) μmol/g/min and (0.66 ± 0.12) μmol/g/min for up to 4 months, comparing well with free, viable cell-specific activities (0.61 ± 0.04 μmol/g/min). Gels cured at 45 °C had excellent structural rigidity and contained few viable cells, making these gels potentially compatible with water treatment facility applications. When encapsulated, non-viable cells were assayed at 4 °C, the activity increased threefold over free cells, potentially due to differences in lipid membranes as shown by FTIR spectroscopy and electron microscopy.

Keywords

Atrazine Silica Bacteria Biodegradation AtzA E. coli 

Supplementary material

253_2011_3821_MOESM1_ESM.doc (74 kb)
ESM 1(DOC 74 kb)

References

  1. Acero JL, Stemmler K, Gunten Uv (2000) Degradation kinetics of atrazine and its degradation products with ozone and OH radicals: a predictive tool for drinking water treatment. Environ Sci Technol 34:591–597CrossRefGoogle Scholar
  2. Acosta EJ, Steffensen MB, Tichy SE, Simanek EE (2004) Removal of water using covalent sequestration. J Agric Food Chem 52:545–549CrossRefGoogle Scholar
  3. Avnir D, Coradin T, Lev O, Livage J (2006) Recent bio-applications of sol–gel materials. J Mater Chem 16:1013–1030CrossRefGoogle Scholar
  4. Bottero JY, Khatib K, Thomas F, Jucker K, Bersillon JL, Mallevialle J (1994) Adsorption of atrazine onto zeolites and organoclays, in the presence of background organics. Water Res 28:483–490CrossRefGoogle Scholar
  5. de Souza ML, Wackett LP, Boundy-Mills KL, Mandelbaum RT, Sadowsky MJ (1995) Cloning, characterization and expression of a gene region from Pseudomonas sp. strain ADP involved in the dechlorination of atrazine. Appl Environ Microbiol 61:3373–3378Google Scholar
  6. Depagne C, Roux C, Coradin T (2011) How to design cell-based biosensors using the sol–gel process. Anal Bioanal Chem 400:965–976CrossRefGoogle Scholar
  7. Devitt E, Wiesner MR (1998) Dialysis investigations of atrazine–organic matter interactions and the role of a divalent metal. Environ Sci Technol 32:232–237CrossRefGoogle Scholar
  8. Doulia D, Hourdakis A, Rigas F, Anagnostopoulus E (1997) Removal of atrazine from water by use of nonionic polymeric resins. J Environ Sci Health, Part A 32:2635–2656Google Scholar
  9. Eisenthal R, Peterson ME, Daniel RM, Danson MJ (2006) The thermal behavior of enzyme activity: implications for biotechnology. Trends Biotechnol 24:289–292CrossRefGoogle Scholar
  10. Ferrer ML, Yuste L, Rojo F, del Monte F (2003) Biocompatible sol–gel route for encapsulation of living bacteria in organically modified silica matrices. Chem Mater 15:3614–3618CrossRefGoogle Scholar
  11. Goswami K, Green RE (1971) Microbial degradation of the herbicide atrazine and its 2-hydroxyanalog in submerged soils. Environ Sci Technol 5:426–429CrossRefGoogle Scholar
  12. Govantes F, Porrúa O, García-González V, Santero E (2009) Atrazine biodegradation in the lab and in the field: enzymatic activities and gene regulation. J Microbiol Biotechnol 2:178–185CrossRefGoogle Scholar
  13. Hosticka B, Norris PM, Brenizer JS, Daitch CE (1998) Gas flow through aerogels. J Non-Cryst Solids 225:293–297CrossRefGoogle Scholar
  14. Kauffmann C, Mandelbaum RT (1998) Entrapment of atrazine chlorohydrolase in sol–gel glass matrix. J Biotechnol 62:168–176CrossRefGoogle Scholar
  15. Le Baron HM, McFarland JE, Burnside OC (2008) The Triazine Herbicides: 50 Years Revolutionizing Agriculture. Elsevier, Oxford, UKGoogle Scholar
  16. Lima D, Viana P, Andre S, Chelinho S, Costa C, Ribeiro R, Sousa JP (2009) Evaluating a bioremediation tool for atrazine contaminated soils in open soil microcosms: the effectiveness of bioaugmentation and biostimulation approaches. Chemosphere 74:187–192CrossRefGoogle Scholar
  17. Lin CH, Lerch RN, Kremer RJ, Garret HE (2011) Stimulated rhizodegradation of atrazine by selected plant species. J Environ Qual 40:1113–1121CrossRefGoogle Scholar
  18. Livage J, Coradin T, Roux C (2001) Encapsulation of biomolecules in silica gels. J Phys Condens Matter 13:R673–R691CrossRefGoogle Scholar
  19. Macias-Flores A, Tafoya-Garnica A, Ruiz-Ordaz N, Salmeron-Alcocer A, Juarez-Ramirez C, Ahuatzi-Chacon D, Mondragon-Parada ME, Galindez-Mayer J (2009) Atrazine biodegradation by a bacterial community immobilized in two types of packed-bed biofilm reactors. World J Microbiol Biotechnol 25:1995–2204CrossRefGoogle Scholar
  20. Majewska-Nowak K, Kabsch-Korutowicz M, Dodz M, Winnicki T (2002) The influence of organic carbon concentration on atrazine removal by UF membranes. Desalination 147:177–122CrossRefGoogle Scholar
  21. Mandelbaum RT, Wackett LP, Allan DL (1993) Rapid hydrolysis of atrazine to hydroxyatrazine by soil bacteria. Environ Sci Technol 27:1943–1946CrossRefGoogle Scholar
  22. Mandelbaum RT, Allan DL, Wackett LP (1995) Isolation and characterization of a Pseudomonas sp. that mineralizes the s-triazine herbicide atrazine. Appl Environ Microbiol 61:1451–1457Google Scholar
  23. Martinez B, Tomkins J, Wackett LP, Wing R, Sadowsky M (2001) Complete nucleotide sequence and organization of the atrazine catabolic plasmid pADP-1 from Pseudomonas sp. strain ADP. J Bacteriol 183:5684–5697CrossRefGoogle Scholar
  24. Nassif N, Bouvet O, Rager MN, Roux C, Coradin T, Livage J (2002) Living bacteria in silica gels. Nat Mater 1:42–44CrossRefGoogle Scholar
  25. Nassif N, Roux C, Coradin T, Bouvet OMM, Livage J (2004) Bacteria quorum sensing in silica matrix. J Mater Chem 14:2264–2268CrossRefGoogle Scholar
  26. Parra S, Stanca SE, Guasaquillo I, Thampi KR (2004) Photocatalytic degradation of atrazine using suspended and supported TiO2. Appl Catal, B 51:107–116CrossRefGoogle Scholar
  27. Pelekani C, Snoeyink VL (2000) Competitive adsorption between atrazine and methylene blue on activated carbon: the importance of pore size distribution. Carbon 38:1423–1436CrossRefGoogle Scholar
  28. Perullini M, Amoura M, Roux C, Coradin T, Livage J, Japas ML, Jobbagy M, Bilmes SA (2011) Improving silica matrices for encapsulation of Escherichia coli using osmoprotectors. J Mater Chem 21:4546–4552CrossRefGoogle Scholar
  29. Reátegui E, Aksan A (2009) Effects of the low temperature transitions of confined water on the structure of isolated and cytoplasmic proteins. J Phys Chem B 113:13048–13060CrossRefGoogle Scholar
  30. Reátegui E, Aksan A (2010) Effects of water on the structure and low/high temperature stability of confined proteins. Phys Chem Chem Phys 12:10161–10172CrossRefGoogle Scholar
  31. Rietti-Shati M, Ronen D, Mandelbaum RT (1996) Atrazine degradation by Pseudomonas strain ADP entrapped in sol–gel glass. J Sol-Gel Sci Technol 7:77–79CrossRefGoogle Scholar
  32. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning; A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  33. Scott C, Lewis SE, Milla R, Taylor MC, Rodgers AJW, Dumsday G, Brodie JE, Oakeshott JG, Russell RJ (2010) A free-enzyme catalyst for the bioremediation of environmental atrazine contamination. J Environ Manag 91:2075–2078CrossRefGoogle Scholar
  34. Seffernick JL, Reynolds E, Fedorov AA, Fedorov E, Almo SC, Sadowsky MJ, Wackett LP (2010) X-ray structure and mutational analysis of the atrazine chlorohydrolase TrzN. J Biol Chem 285:30606–30614CrossRefGoogle Scholar
  35. Silva E, Fialho AM, Sá-Correia I, Burns RG, Shaw LJ (2004) Combined bioaugmentation and biostimulation to cleanup soil contaminated with high concentrations of atrazine. Environ Sci Technol 38:632–637CrossRefGoogle Scholar
  36. Strong LC, McTavish H, Sadowsky MJ, Wackett LP (2000) Field-scale remediation of atrazine-contaminated soil using recombinant Escherichia coli expressing atrazine chlorohydrolase. Environ Microbiol 2:91–98CrossRefGoogle Scholar
  37. Topp E, Mulbry WM, Zhu H, Nour SM, Cuppels D (2000) Characterization of s-triazine herbicide metabolism by a Nocardioides sp. isolated from agricultural soils. Appl Environ Microbiol 66:3134–3141CrossRefGoogle Scholar
  38. Wang L, Samac DA, Shapir N, Wackett LP, Vance CP, Olszewski NE, Sadowsky MJ (2005) Biodegradation of atrazine in transgenic plants expressing a modified bacterial atrazine chlorohydrolase (atzA) gene. J Plant Biotechnol 3:475–486CrossRefGoogle Scholar
  39. Yue Z, Economy J, Rajagopalan K, Bordson G, Piwoni M, Ding L, Snoeyink VL, Marinas BJ (2006) Chemical activated carbon on a fiberglass substrate for removal of trace atrazine from water. J Mater Chem 16:3375–3380CrossRefGoogle Scholar
  40. Zhu L, Ma T, Wang J, Xie H, Wang J, Xin C, Shao B (2011) Enhancement of atrazine removal by free and immobilized Arthrobacter sp. HB-5 in soil and wastewater. Soil Sediment Contam 20:87–97CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Eduardo Reátegui
    • 1
  • Erik Reynolds
    • 4
  • Lisa Kasinkas
    • 1
  • Amit Aggarwal
    • 3
  • Michael J. Sadowsky
    • 4
    • 5
  • Alptekin Aksan
    • 1
    • 4
  • Lawrence P. Wackett
    • 2
    • 4
  1. 1.Biostabilization Laboratory, Department of Mechanical EngineeringUniversity of MinnesotaMinneapolisUSA
  2. 2.Department of Biochemistry, Molecular Biology and BiophysicsUniversity of MinnesotaMinneapolisUSA
  3. 3.Department of Bioproducts and Biosystems EngineeringUniversity of MinnesotaSaint PaulUSA
  4. 4.BioTechnology InstituteUniversity of MinnesotaSaint PaulUSA
  5. 5.Department of Soil, Water, and ClimateUniversity of MinnesotaSaint PaulUSA

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