Issue

Vol. 28 No. 11 (2016): Vol 28 Issue 11

Copyright (c) 2016 AJC

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Accelerated Production of Oxygen-Insensitive Azoreductase from Mutant Pseudomonas Species for Degradation Azo Dyes under Aerobic Condition

https://doi.org/10.14233/ajchem.2016.20113
Arunkumar Mani
Department of Chemistry & Biosciences, Srinivasa Ramanujan Centre, SASTRA University, Kumbakonam-612 001, India
Sheik Abdulla Shahul Hameed
Department of Chemistry & Biosciences, Srinivasa Ramanujan Centre, SASTRA University, Kumbakonam-612 001, India

Corresponding Author(s) : Arunkumar Mani

arunkumar@src.sastra.edu; microarun2008@yahoo.in

Asian Journal of Chemistry, Vol. 28 No. 11 (2016): Vol 28 Issue 11

Abstract

Bacteria producing an oxygen insensitive intracellular azoreductase were isolated from a dye contaminated area and identified as a Pseudomonas species B1 by 16s rRNA sequencing. Maximum azoreductase production (0.39 U/mL) was observed in the mutant strain within 30 h of incubation under optimized conditions at pH 6.5, temperature 35 °C, glucose 2 %, sodium nitrate 1 % and 2 % of inoculum concentration. Azoreductase was purified by ammonium precipitation method and followed by anion exchange chromatography. Mutant Pseudomonas species B1 showed 2-fold increased level of azoreductase production and purified up to 94-fold with a recovery of 18 %. Native PAGE analysis revealed that the purified enzyme was a monomer with a molecular weight of 29 kDa. The Km and Vmax values were 0.09 mM and 6.7 U mg–1 of protein for NADH and 0.04 mM and 4.7 U mg–1 of protein for naphthol blue black, respectively. Furthermore, the purified enzyme could effectively degrade 78 % of naphthol blue black under aerobic conditions, as monitored by UV-visible, FTIR-spectroscopy, HPLC and GCMS. Phytotoxicity and microbial toxicity assays showed that the degradation products of naphthol blue black were less toxic than the dye itself.

Keywords

Azoreductase Pseudomonas sp. Random mutagenesis Dye decolorization Phytotoxicity
Mani, A., & Shahul Hameed, S. A. (2016). Accelerated Production of Oxygen-Insensitive Azoreductase from Mutant Pseudomonas Species for Degradation Azo Dyes under Aerobic Condition. Asian Journal of Chemistry, 28(11), 2562–2570. https://doi.org/10.14233/ajchem.2016.20113
Download Citation
Endnote/Zotero/Mendeley (RIS)
BibTeX
References
  1. K.T. Chung Jr., S.E. Stevens and C.E. Cerniglia, Crit. Rev. Microbiol., 18, 175 (1992); doi:10.3109/10408419209114557.
  2. A. Stolz, Appl. Microbiol. Biotechnol., 56, 69 (2001); doi:10.1007/s002530100686.
  3. H. Chen, Curr. Protein Pept. Sci., 7, 101 (2006); doi:10.2174/138920306776359786.
  4. J. Feng, T.M. Heinze, H. Xu, C.E. Cerniglia and H. Chen, Protein Pept. Lett., 17, 578 (2010); doi:10.2174/092986610791112701.
  5. R. Anliker, Ecotoxicol. Environ. Saf., 3, 59 (1979); doi:10.1016/0147-6513(79)90060-5.
  6. D. Cui, G. Li, D. Zhao, X. Gu, C. Wang and M. Zhao, J. Hazard. Mater., 221-222, 185 (2012); doi:10.1016/j.jhazmat.2012.04.032.
  7. H.S. Lade, T.R. Waghmode, A.A. Kadam and S.P. Govindwar, Int. Biodeter. Biodegrad., 72, 94 (2012); doi:10.1016/j.ibiod.2012.06.001.
  8. A.A. Telke, D.C. Kalyani, V.V. Dawkar and S.P. Govindwar, J. Hazard. Mater., 172, 298 (2009); doi:10.1016/j.jhazmat.2009.07.008.
  9. A. Ryan, N. Laurieri, I. Westwood, C.J. Wang, E. Lowe and E. Sim, J. Mol. Biol., 400, 24 (2010); doi:10.1016/j.jmb.2010.04.023.
  10. W. Lang, S. Sirisansaneeyakul, L. Ngiwsara, S. Mendes, L.O. Martins, M. Okuyama and A. Kimura, Bioresour. Technol., 150, 298 (2013); doi:10.1016/j.biortech.2013.09.124.
  11. J. Feng, O. Kweon, H. Xu, C.E. Cerniglia and H. Chen, Arch. Biochem. Biophys., 520, 99 (2012); doi:10.1016/j.abb.2012.02.010.
  12. K. Tamura, D. Peterson, N. Peterson, G. Stecher, M. Nei and S. Kumar, Mol. Biol. Evol., 28, 2731 (2011); doi:10.1093/molbev/msr121.
  13. N. Pfennig, Arch. Microbiol., 100, 197 (1974); doi:10.1007/BF00446317.
  14. C.V. Nachiyar and G.S. Rajakumar, Enzyme Microb. Technol., 36, 503 (2005); doi:10.1016/j.enzmictec.2004.11.015.
  15. O.H. Lowry, N.J. Rosebrough, A.L. Farr and R.J. Randall‎, J. Biochem., 193, 265 (1951).
  16. S.A. Misal, D.P. Lingojwar, R.M. Shinde and K.R. Gawai, Process Biochem., 46, 1264 (2011); doi:10.1016/j.procbio.2011.02.013.
  17. R.G. Saratale, G.D. Saratale, J.S. Chang and S.P. Govindwar, Bioresour. Technol., 100, 3897 (2009); doi:10.1016/j.biortech.2009.03.051.
  18. S. Nouren and H.N. Bhatti, Biochem. Eng., 95, 9 (2015); doi:10.1016/j.bej.2014.11.021.
  19. N. Saitou and M. Nei, Mol. Biol. Evol., 4, 406 (1987).
  20. K. Tamura, M. Nei and S. Kumar, Proc. Natl. Acad. Sci. USA, 101, 11030 (2004); doi:10.1073/pnas.0404206101.
  21. S. Dhawan, R. Lal and R.C. Kuhad, Lett. Appl. Microbiol., 36, 64 (2003); doi:10.1046/j.1472-765X.2003.01267.x.
  22. B. Naveena, K.P. Gopinath, P. Sakthiselvan and N. Partha, Bioresour. Technol., 111, 417 (2012); doi:10.1016/j.biortech.2012.02.056.
  23. T. Zimmermann, F. Gasser, H.G. Kulla and T. Leisinger, Arch. Microbiol., 138, 37 (1984); doi:10.1007/BF00425404.
  24. D.K. Ghosh, S. Ghosh, P. Sadhukhan, A. Mandal and J. Chauduri, Indian J. Exp. Biol., 31, 951 (1993).
  25. T.L. Hu, Water Sci. Technol., 43, 261 (2001).
  26. J. Maier, A. Kandelbauer, A. Erlacher, A. Cavaco-Paulo and M. Gubitz, Appl. Environ. Microbiol., 70, 837 (2004); doi:10.1128/AEM.70.2.837-844.2004.
  27. K.P. Gopinath, S. Murugesan, J. Abraham and K. Muthukumar, Bioresour. Technol., 100, 6295 (2009); doi:10.1016/j.biortech.2009.07.043.
  28. E. Torres, I. Bustos-Jaimes and S. Le Borgne, Appl. Catal. B, 46, 1 (2003); doi:10.1016/S0926-3373(03)00228-5.
  29. M.S. Kim, D.H. Kim and J. Cha, Int. J. Hydrogen Energy, 37, 14055 (2012); doi:10.1016/j.ijhydene.2012.06.085.
  30. R. Mazumder, J.R. Logan, A.T. Mikell Jr and S.W. Hooper, Industrial Microbiol. Biotechnol., 23, 476 (1999); doi:10.1038/sj.jim.2900734.
  31. G. Liu, J. Zhou, J. Wang, M. Zhou, H. Lu and R. Jin, Bioresour. Technol., 100, 2791 (2009); doi:10.1016/j.biortech.2008.12.040.
  32. R. Russ, J. Rau and A. Stolz, Appl. Environ. Microbiol., 66, 1429 (2000); doi:10.1128/AEM.66.4.1429-1434.2000.
  33. S. Blumel, H.J. Knackmuss and A. Stolz, Appl. Environ. Microbiol., 68, 3948 (2002); doi:10.1128/AEM.68.8.3948-3955.2002.
  34. E. Idaka, T. Ogawa and H. Horitsu, Environ. Contam. Toxicol., 39, 108 (1987); doi:10.1007/BF01691797.
  35. S.W. Hooper, Proteases in Biology and Medicine, In: Essays in Biochemistry, Portland Press, vol. 25, pp. 169-182 (1994).
  36. M. Nakanishi, C. Yatome, N. Ishida and Y. Kitade, J. Biol. Chem., 276, 46394 (2001); doi:10.1074/jbc.M104483200.
Read More
Author biographies is not available.
Download
PDF
Statistic
Read Counter : 0 Download : 0