Skip to main content

Isolation and identification of Pseudomonas azotoformans for induced calcite precipitation

World Journal of Microbiology and Biotechnology volume 31, pages 1993–2001 (2015)Cite this article

Abstract

Biomineralization is a process by which living organisms produce minerals. The extracellular production of these biominerals by microbes has potential for various bioengineering applications. For example, crack remediation and improvement of durability of concrete is an important goal for engineers and biomineral-producing microbes could be a useful tool in achieving this goal. Here we report the isolation, biochemical characterization and molecular identification of Pseudomonas azotoformans, a microbe that produces calcite and which potentially be used to repair cracks in concrete structures. Initially, 38 bacterial isolates were isolated from soil and cements. As a first test, the isolates were screened using a urease assay followed by biochemical tests for the rate of urea hydrolysis, calcite production and the insolubility of calcite. Molecular amplification and sequencing of a 16S rRNA fragment of selected isolates permitted us to identify P. azotoformans as a good candidate for preparation of biotechnological concrete. This species was isolated from soil and the results show that among the tested isolates it had the highest rate of urea hydrolysis, produced the highest amount of calcite, which, furthermore was the most adhesive and insoluble. This species is thus of interest as an agent with the potential ability to repair cracks in concrete.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  • Abo-El-Enein SA, Ali AH, Talkhan FN, Abdel-Gawwad HA (2012) Utilization of microbial induced calcite precipitation for sand consolidation and mortar crack remediation. HBRC 8:185–192

    Article  Google Scholar 

  • Achal V, Pan X (2014) Influence of calcium sources on microbially induced calcium carbonate precipitation by Bacillus sp. CR2. Appl Biochem Biotechnol 173:307–317

    CAS  Article  Google Scholar 

  • Achal V, Mukherjee A, Goyal Sh, Reddy S (2012) Corrosion prevention of reinforced concrete with microbial calcite precipitation. ACI Mater J 2:157–164

    Google Scholar 

  • Anzai Y, Kim H, Park JY, Wakabayashi H, Oyaizu H (2000) Phylogenetic affiliation of the Pseudomonads based on 16S rRNA sequence. Int J Syst Evol Microbiol 50:1563–1589

    CAS  Article  Google Scholar 

  • Bachmeier KL, Williams AE, Warmington JR, Bang SS (2002) Urease activity in microbiologically-induced calcite precipitation. J Biotechnol 93:171–181

    CAS  Article  Google Scholar 

  • Bang SS, Galinat JK, Ramakrishnan V (2001) Calcite precipitation induced by polyurethane-immobilized Bacillus pasteurii. Enzyme Microb Technol 28:404–409

    CAS  Article  Google Scholar 

  • Chahal N, Rajor A, Siddique R (2011) Calcium carbonate precipitation by different bacterial strains. Afr J Biotechnol 10:8359–8372

    CAS  Google Scholar 

  • De Belie N, De Muynck W (2009) Crack repair in concrete using biodeposition. In: Alexander MG, Beushausen H-D, Dehn F, Moyo P (eds) Concret repair, rehabilitation and retrofitting II. Taylor and Francis Group, London, pp 777–781

    Google Scholar 

  • De Belie N, De Muynck W, Verstraete W (2006) A synergistic approach to microbial presence on concrete: cleaning and consolidating effects. Struct Concr 7:105–109

    Article  Google Scholar 

  • De Muynck W, De Belie N, Verstraete W (2007). Improvement of concrete durability with the aid of bacteria. In: Proceedings of the 1st international conference on self-healing materials, Noordwijk aan Zee, The Netherlands, April 2007, pp 18–20

  • DeJong JT, Mortensen BM, Martinez BC, Nelson DC (2010) Bio-mediated soil improvement. Ecol Eng 36:197–210

    Article  Google Scholar 

  • Dhami NK, Reddy MS, Mukherjee A (2013a) Biomineralization of calcium carbonates and their engineered applications: a review. Front Microbiol 4:314. doi:10.3389/fmicb.2013.00314

    Google Scholar 

  • Dhami NK, Reddy MS, Mukherjee A (2013b) Biomineralization of calcium carbonate polymorphs by the bacterial strains isolated from calcareous sites. J Microbiol Biotechnol 23:707–714

    CAS  Article  Google Scholar 

  • Dick J, De Windt W, De Graef B, Saveyn H, Van der Meeren P, De Belie N, Verstraete W (2006) Bio-deposition of a calcium carbonate layer on degraded limestone by Bacillus species. Biodegradation 17:357–367

    CAS  Article  Google Scholar 

  • Ferris FG, Stehmeier LG (1992). Bacteriogenic mineral plugging. USA Patent 5,143, 155. US Patent Office, Washington, DC

  • Ghosh P, Mandal S, Chattopadhyay BD, Pal S (2005) Use of microorganism to improve the strength of cement mortar. Cem Concr Res 35:1980–1983

    CAS  Article  Google Scholar 

  • Gollapudi UK, Knutson CL, Bang SS, Islam MR (1995) A new method for controlling leaching through permeable channels. Chemosphere 30:695–705

    CAS  Article  Google Scholar 

  • Hammes F, Boon N, de Villiers J, Verstraete W, Siciliano SD (2003) Strain-specific ureolytic microbial calcium carbonate precipitation. J Appl Environ Microbiol 69:4901–4909

    CAS  Article  Google Scholar 

  • Ivanov V, Chu J (2008) Applications of microorganisms to geotechnical engineering for bioclogging and biocementation of soil in situ. Rev Environ Sci Biotechnol 7:139–153

    CAS  Article  Google Scholar 

  • Jagadeesha-Kumar BG, Prabhakara R, Pushpa H (2013) Bio Mineralisation of calcium carbonate by diferent bacterial strains and their application in concrete crack remediation. Int J Adv Eng Technol 6:202–213

    Google Scholar 

  • Jimenez-Lopez C, Rodriguez-Navarro C, Piñar G, Carrillo-Rosúa FJ, Rodriguez-Gallego M, Gonzalez-Muñoz MT (2008) Consolidation of quarry calcarenite by calcium carbonate precipitation induced by bacteria activated among the microbiota inhabiting the stone. Int Biodeterior Biodegrad 62:352–363

    CAS  Article  Google Scholar 

  • Jonkers HM, Thijssen A, Muyzer G, Copuroglu O et al (2010) Application of bacteria as self-healing agent for the development of sustainable concrete. Ecol Eng 36:230–235

    Article  Google Scholar 

  • Klement Z, Rudolph K, Sands DC (1990) Methods in phytobacteriology. Akademiai Kiado Press, Budapest, pp 133–143

    Google Scholar 

  • Le Metayer-Levrel G, Castanier S, Orial G, Loubiere JF et al (1999) Applications of bacterial carbonatogenesis to the protection and regeneration of limestones in buildings and historic patrimony. Sediment Geol 126:25–34

    Article  Google Scholar 

  • Lopez-Moreno A, Sepulveda-Sanchez JD, Mercedes Alonso Guzman EM, Le Borgne S (2014) Calcium carbonate precipitation by heterotrophic bacteria isolated from biofilms formed on deteriorated ignimbrite stones: influence of calcium on EPS production and biofilm formation by these isolates. Biofouling 30:547–560

    CAS  Article  Google Scholar 

  • Lowenstam HA (1981) Minerals formed by organisms. Science 211:1126–1131

    CAS  Article  Google Scholar 

  • Nemati M, Voordouw G (2003) Modification of porous media permeability using calcium carbonate produced enzymatically in situ. Enzyme Microb Technol 33:635–642

    CAS  Article  Google Scholar 

  • Okwadha G, Li J (2010) Optimum conditions for microbial carbonate precipitation. Chemosphere 81:1143–1148

    CAS  Article  Google Scholar 

  • Phillips AJ, Gerlach R, Lauchnor E, Mitchell AC, Cunningham AB, Spangler L (2013) Engineered applications of ureolytic biomineralization: a review. Biofouling 29:715–733

    CAS  Article  Google Scholar 

  • Raut SH, Sarode DD, Lele SS (2014) Biocalcification using B. pasteurii for strengthening brick masonry civil engineering structures. World J Microbiol Biotechnol 30:191–200

    CAS  Article  Google Scholar 

  • Rivadeneyra MA, Párraga J, Delgado R, Ramos-Cormenzana A, Delgado G (2004) Biomineralization of carbonates by Halobacillus trueperi in solid and liquid media with different salinities. FEMS Microbiol Ecol 48:39–46

    CAS  Article  Google Scholar 

  • Rodriguez-Navarro C, Rodriguez-Gallego M, Chekroun KB, Gonzalez-Munoz MT (2003) Conservation of ornamental stone by Myxococcus xanthus-induced carbonate biomineralization. Appl Environ Microbiol 69:2182–2193

    CAS  Article  Google Scholar 

  • Sarayu K, Iyer NR, Murthy AR (2014) Exploration on the biotechnological aspect of the ureolytic bacteria for the production of the cementitious materials—a review. Appl Biochem Biotechnol 172:2308–2323

    CAS  Article  Google Scholar 

  • Stocks-Fischer S, Galinat JK, Bang SS (1999) Microbiological precipitation of CaCO3. Soil Biol Biochem 31:1563–1571

    CAS  Article  Google Scholar 

  • Tiano P, Biagiotti L, Mastromei G (1999) Bacterially bio-mediated calcite precipitation for monumental stones conservation: methods of evaluation. J Microbiol Methods 36:139–145

    CAS  Article  Google Scholar 

  • Tittelboom KV, Nele K, De Belie N, De Muynck W et al (2010) Use of bacteria to repair cracks in concrete. Cem Concr Res 40:157–166

    Article  Google Scholar 

  • Vahabi A, Ramezanianpour AA, Sharafi H, Zahiri HS, Vali H, Noghabi KA (2015) Calcium carbonate precipitation by strain Bacillus licheniformis AK01, newly isolated from loamy soil: a promising alternative for sealing cement-based materials. J Basic Microbiol 55:105–111

    CAS  Article  Google Scholar 

  • Wang Y, Qian PY (2009) Conservative fragments in bacterial 16S rRNA genes and primer design for 16S ribosomal DNA amplicons in metagenomic studies. PLoS ONE 4:e7401

    Article  Google Scholar 

  • Whiffin VS, Van Paassen LA, Harkes MP (2007) Microbial carbonate precipitation as a soil improvement technique. Geomicrobiol J 24:417–423

    CAS  Article  Google Scholar 

Download references

Acknowledgments

This research has been supported by Azarbaijan Shahid Madani University. Special thanks to IBMP-CNRS Strasbourg, France for DNA sequencing and very special thanks to Kenneth E. Richards for English language revising.

Author information

Affiliations

  1. Faculty of Engineering, Azarbijan Shahid Madani University, Tabriz, Iran

    Siamak Heidari Nonakaran & Abdullah Keyvani

  2. Biotechnology Department, Agriculture Faculty, Azarbijan Shahid Madani University, Km 35 Tabriz-Azarshahr Road, Tabriz, Iran

    Maghsoud Pazhouhandeh, Fatemeh Zahra Abdollahipour & Akbar Shirzad

Authors
  1. Siamak Heidari Nonakaran
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maghsoud Pazhouhandeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Abdullah Keyvani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fatemeh Zahra Abdollahipour
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Akbar Shirzad
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maghsoud Pazhouhandeh.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Heidari Nonakaran, S., Pazhouhandeh, M., Keyvani, A. et al. Isolation and identification of Pseudomonas azotoformans for induced calcite precipitation. World J Microbiol Biotechnol 31, 1993–2001 (2015). https://doi.org/10.1007/s11274-015-1948-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-015-1948-5

Keywords

  • Pseudomonas
  • Crack remediation
  • Calcite production
  • Concrete