Advertisement

Annals of Microbiology

, Volume 68, Issue 12, pp 931–942 | Cite as

Immobilization of Pseudomonas putida PT in resistant matrices to environmental stresses: a strategy for continuous removal of heavy metals under extreme conditions

  • Sanaz Khashei
  • Zahra EtemadifarEmail author
  • Hamid Reza Rahmani
Original Article
  • 64 Downloads

Abstract

The purpose of this study was to investigate the potential of immobilized lead- and cadmium-resistant Pseudomonas putida strain PT to remove heavy metals from aqueous medium under extreme conditions. The tolerance and accumulation of cadmium and lead ions by strain PT were investigated by minimal inhibitory concentration (MIC) determination and polymerase chain reaction (PCR) of cadA gene, respectively. The surface chemical functional groups of P. putida PT involved in the metal biosorption were identified by Fourier transform infrared (FTIR). Pseudomonas putida PT was immobilized in three matrices include carboxy-methyl cellulose (CMC), rice bran, and a new composite made of alginate, polyvinyl alcohol (PVA), and CaCO3 to prepare heavy metal adsorbent. The biosorbents were analyzed by SEM, and their metal removal capability was assayed in two consecutive cycles by atomic absorption spectroscopy. The viability of immobilized bacterial cells was determined by flow cytometry during storage at 4 °C and exposure to the environmental stresses (pH and temperature). The results showed that PT strain was resistant up to 10 mM Pb2+ and 8 mM Cd2+. FTIR analysis revealed that alcohol, sulfur, phosphate, esters, and amide groups played important roles in metal biosorption process and, also change in metabolic reactions like hydration and polyesters accumulation was observed after metal biosorption. The presence of cadA gene, a heavy metal translocating pump-coding gene, indicated the ability of metals bioaccumulation by the PT strain. Immobilized cells in alginate–PVA–CaCO3 and rice bran showed the highest metal removal efficiency for Pb2+ as 75% and Cd2+ as 96.7%, respectively. Metal adsorbents were reusable, and the highest removal efficiency in the second cycle was observed in inoculated alginate–PVA–CaCO3 (79.5% Pb2+ and 45% Cd2+). Flow cytometric analysis represented that the immobilized cell viability was retained (< 97%) after 4 weeks storage at 4 °C. Viability under two environmental stresses in all matrices was as follows: < 96% at 25 °C, < 87% at 45 °C, < 85% at pH 4, < 96% at pH 7, and < 89% at pH 11. The results signify that these metal adsorbents are efficient technological tools for bioremediation even in harsh environmental conditions.

Keywords

Heavy metals Biosorption Bioaccumulation Microbe immobilization Viability 

Notes

Funding

This study was funded by the University of Isfahan and Soil and Water Research Institute of Iran.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

13213_2018_1402_MOESM1_ESM.jpg (683 kb)
Fig. A Viability of immobilized P. putida PT during storage at 4 °C. Bacterial cell samples were stained with Rho - 123 and analyzed by flow cytometer (FL1, 525 nm). a, b, c and d indicate the viability of immobilized bacterial cells on rice bran, e, f, g and h represent the viability of immobilized bacterial cells in CMC and i, j, k and l show the viability of immobilized bacterial cells in alginate-PVA-CaCO3 from week 1 to week 4 (JPG 683 kb)
13213_2018_1402_MOESM2_ESM.jpg (622 kb)
Fig. B Flow cytometric analysis of P. putida PT at different pHs. a, b and c indicate the viability of immobilized bacterial cells on rice bran, e, f and g represent the viability of immobilized bacterial cells in CMC and i, j and k show the viability of immobilized bacterial cells in alginate-PVA-CaCO3 at pH 4, 7 and 10, respectively (JPG 621 kb)
13213_2018_1402_MOESM3_ESM.jpg (430 kb)
Fig. C Flow cytometric analysis of P. putida PT at different temperatures. a and d indicate the viability of immobilized bacterial cells on rice bran, b and e represent the viability of immobilized bacterial cells in CMC and c and f show the viability of immobilized bacterial cells in alginate-PVA-CaCO3 at 25 °C and 45 °C, respectively (JPG 429 kb)

References

  1. Ahmaruzzaman M, Gupta VK (2011) Rice husk and its ash as low-cost adsorbents in water and wastewater treatment. Ind Eng Chem Res 50:13589–13613CrossRefGoogle Scholar
  2. Baatout S, De Boever P, Mergeay M (2006) Physiological changes induced in four bacterial strains following oxidative stress. Appl Biochem Microbiol 42:369–377CrossRefGoogle Scholar
  3. Bashan Y, de-Bashan LE, Prabhu S, Hernandez J-P (2014) Advances in plant growth-promoting bacterial inoculant technology: formulations and practical perspectives (1998–2013). Plant Soil 378:1–33CrossRefGoogle Scholar
  4. Bayat Z, Hassanshahian M, Cappello S (2015) Immobilization of microbes for bioremediation of crude oil polluted environments: a mini review. Open Microbiol J 9:48PubMedPubMedCentralGoogle Scholar
  5. Berninger T, González López Ó, Bejarano A, Preininger C, Sessitsch A (2018) Maintenance and assessment of cell viability in formulation of non-sporulating bacterial inoculants. Microb Biotechnol 11:277–301CrossRefGoogle Scholar
  6. Bi C, Zhou Y, Chen Z, Jia J, Bao X (2018) Heavy metals and lead isotopes in soils, road dust and leafy vegetables and health risks via vegetable consumption in the industrial areas of Shanghai, China. Sci Total Environ 619:1349–1357CrossRefGoogle Scholar
  7. Chandran P, Das N (2011) Degradation of diesel oil by immobilized Candida tropicalis and biofilm formed on gravels. Biodegradation 22:1181–1189CrossRefGoogle Scholar
  8. Chen L, Zhou S, Shi Y, Wang C, Li B, Li Y, Wu S (2018) Heavy metals in food crops, soil, and water in the Lihe River Watershed of the Taihu Region and their potential health risks when ingested. Sci Total Environ 615:141–149CrossRefGoogle Scholar
  9. Cruz I, Bashan Y, Hernàndez-Carmona G, De-Bashan LE (2013) Biological deterioration of alginate beads containing immobilized microalgae and bacteria during tertiary wastewater treatment. Appl Microbiol Biotechnol 97:9847–9858CrossRefGoogle Scholar
  10. Das M, Adholeya A (2015) Potential uses of immobilized bacteria, fungi, algae, and their aggregates for treatment of organic and inorganic pollutants in wastewater. In: Water challenges and solutions on a global scale. ACS Publications, pp 319–337Google Scholar
  11. Giti E, Mehdi H, Nasser G (2005) Development of a microtitre plate method for determination of phenol utilization, biofilm formation and respiratory activity by environmental bacterial isolates. Int Biodeterior Biodegrad 56:231–235CrossRefGoogle Scholar
  12. Hazaimeh M, Mutalib SA, Abdullah PS, Kee WK, Surif S (2014) Enhanced crude oil hydrocarbon degradation by self-immobilized bacterial consortium culture on sawdust and oil palm empty fruit bunch. Ann Microbiol 64:1769–1777CrossRefGoogle Scholar
  13. Hedlund B, Yoon J, Kasai H (2015) Bergey’s manual of systematics of archaea and bacteria. Wiley, HobokenGoogle Scholar
  14. Hynninen A (2010) Zinc, cadmium and lead resistance mechanisms in bacteria and their contribution to biosensing. Dissertation, University of HelsinkiGoogle Scholar
  15. Icgen B, Yilmaz F (2016) Design a cadA-targeted DNA probe for screening of potential bacterial cadmium biosorbents. Environ Sci Pollut Res Int 23:5743–5752CrossRefGoogle Scholar
  16. Krishnani KK, Meng X, Christodoulatos C, Boddu VM (2008) Biosorption mechanism of nine different heavy metals onto biomatrix from rice husk. J Hazard Mater 153:1222–1234CrossRefGoogle Scholar
  17. Liaqat I (2017) Heavy metal bioremediation in soil: key species and strategies involved in the process. IJABF 1:38–48Google Scholar
  18. Maitra S (2016) Study of genetic determinants of nickel and cadmium resistance in bacteria—a review. Int J Curr Microbiol App Sci 5:459–471CrossRefGoogle Scholar
  19. Marzan LW, Hossain M, Mina SA, Akter Y, Chowdhury AMA (2017) Isolation and biochemical characterization of heavy-metal resistant bacteria from tannery effluent in Chittagong city, Bangladesh: bioremediation viewpoint. Egypt J Aquat Res 43:65–74CrossRefGoogle Scholar
  20. Naz N, Young HK, Ahmed N, Gadd GM (2005) Cadmium accumulation and DNA homology with metal resistance genes in sulfate-reducing bacteria. Appl Environ Microbiol 71:4610–4618CrossRefGoogle Scholar
  21. Nies DH (2003) Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 27:313–339CrossRefGoogle Scholar
  22. Oves M, Zaidi A, Khan MS (2010) Role of metal tolerant microbes in legume improvement. In: Microbes for legume improvement. Springer, Berlin, pp 337–352CrossRefGoogle Scholar
  23. Oves M, Khan MS, Zaidi A (2013) Biosorption of heavy metals by Bacillus thuringiensis strain OSM29 originating from industrial effluent contaminated north Indian soil. Saudi J Biol Sci 20:121–129CrossRefGoogle Scholar
  24. Özdemir S, Kilinc E, Poli A, Nicolaus B, Güven K (2012) Cd, Cu, Ni, Mn and Zn resistance and bioaccumulation by thermophilic bacteria, Geobacillus toebii subsp. decanicus and Geobacillus thermoleovorans subsp. stromboliensis. World J Microbiol Biotechnol 28:155–163CrossRefGoogle Scholar
  25. Pal A, Datta S, Paul AK (2013) Hexavalent chromium reduction by immobilized cells of Bacillus sphaericus AND 303. Braz Arch Biol Technol 56:505–512CrossRefGoogle Scholar
  26. Pires C, Marques AP, Guerreiro A, Magan N, Castro PM (2011) Removal of heavy metals using different polymer matrixes as support for bacterial immobilisation. J Hazard Mater 191:277–286CrossRefGoogle Scholar
  27. Rani MJ, Hemambika B, Hemapriya J, Kannan VR (2010) Comparative assessment of heavy metal removal by immobilized and dead bacterial cells: a biosorption approach. AJEST 4:77–83Google Scholar
  28. Shamim S, Rehman A, Qazi MH (2014) Cadmium-resistance mechanism in the bacteria Cupriavidus metallidurans CH34 and Pseudomonas putida mt2. Arch Environ Contam Toxicol 67:149–157CrossRefGoogle Scholar
  29. Tchounwou PB, Yedjou CG, Patlolla AK, Sutton DJ (2012) Heavy metal toxicity and the environment In: Molecular, clinical and environmental toxicology Springer, pp 133–164Google Scholar
  30. Wen X, Du C, Zeng G, Huang D, Zhang J, Yin L, Tan S, Huang L, Chen H, Yu G, Hu X (2018) A novel biosorbent prepared by immobilized Bacillus licheniformis for lead removal from wastewater. Chemosphere 200:173–179CrossRefGoogle Scholar
  31. Wu Z, Guo L, Qin S, Li C (2012) Encapsulation of R. planticola Rs-2 from alginate-starch-bentonite and its controlled release and swelling behavior under simulated soil conditions. J Ind Microbiol Biotechnol 39:317–327CrossRefGoogle Scholar
  32. Yang S, Fu S, Liu H, Zhou Y, Li X (2011) Hydrogel beads based on carboxymethyl cellulose for removal heavy metal ions. J Appl Polym Sci 119:1204–1210CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018

Authors and Affiliations

  1. 1.Department of Biology, Faculty of SciencesUniversity of IsfahanIsfahanIran
  2. 2.Environmental Research InstituteUniversity of IsfahanIsfahanIran
  3. 3.Department of Soil and Water Research, Agricultural Research, Educating and Extension Organization, AREEOIsfahanIran

Personalised recommendations