Decolourisation of Reactive Black 5 by Azoreductase Produced by Brevibacillus panacihumi ZBI
DOI:
https://doi.org/10.11113/jt.v59.1571Keywords:
Azoreductase, brevibacillus panacihumi ZBI, reactive Black 5, decolourisation, azoreductase assayAbstract
Azoreductases are often associated with decolourisation of non–degradable azo dyes via cleavage of azo bonds. In this study,Brevibacillus panacihumi ZBI, an azo dye–degrading bacterium which has not been reported before, was used for the decolourisation of Reactive Black 5 (RB5) dye. The highest activity of azoreductase was obtained during the end of log phase. Azoreductase produced intracellularly had the highest specific activity of 0.334 U/mg compared to the culture supernatant (extracellular), resting cell and cell debris with low enzyme activity of 0.034 U/mg, 0.010 U/mg and 0.200 U/mg respectively. The optimum assay conditions for the maximum azoreductase activity were at 37°C, pH 7, RB5 dye concentration of 100 mg/L and NADH concentration of 0.2 mM by using phosphate buffer as a medium for the enzyme reaction. Alternatively, the azoreductase assay was also carried out using ionic liquid, [emim][EtSO4] that may function to enhance the activity and stability of azoreductase. Results using phosphate buffer (pH 7) showed higher enzyme activity twice that of the ionic liquid besides enhancing the stability of enzyme. Under the optimum assay conditions up to 93 % of decolourisation was achieved after 8 h of incubation. In addition, growth of bacteria was also concurrently observed during the decolourisation of RB5.References
Jin, X. C., Liu, G. Q., Xu, Z. H., and Tao, W. Y. 2007. Decolourization
of a Dye Industry Effluent by Aspergillus fumigates XC6. Applied
Microbiology and Biotechnology. 74: 239–243.
Jadhav, J. P., Kalyani, D. C., Telke, A. A., Phugare, S. S., and
Govindwar, S. P. 2010. Evaluation of the Efficacy of a Bacterial
Consortium for the Removal of Colour, Reduction of Heavy Metals and
Toxicity from Textile Dye Effluent. Bioresources Technology. 101: 165–
Forgacs, E., Cserhati, T., and Oros, G. 2004. Removal of Synthetic Dyes
from Wastewaters. A Review. Environmental International. 30: 953–
Dafale, N., Wate, S., Meshram, S., and Neti, R. N. 2010. Bioremediation
of Wastewater Containing Azo Dyes Through Sequential Anaerobic–
Aerobic Bioreactor System and Its Biodiversity. Environmental Reviews.
: 21–36.
Ghosh, D. K., Mandal, A., and Chaudhuri, J. 1992. Purification and
Partial Characterization of Two Azoreductases from Shigella Dysenteriae
Type1, FEMS Microbiology Letters. 77: 229–233.
Moutaouakkil, A., Zeroual, Y., Dzayri, F. Z., Talbi, M., Lee, K., and
Blaghen, M. 2003. Purification and Partial Characterization of
Azoreductase From Enterobacter agglomerans. Archieves of
Biochemistry and Biophysics. 413: 139–146.
Chen, H., Hopper, S. L., and Cerniglia, C. E. 2005. Biochemical And
Molecular Characterization of an Azoreductase From Staphylococcus
aureus, a tetrameric NADPH-Dependent Flavoprotein. Microbiology.
: 1433–1441.
Olukanni, O. D., Osuntoki, A. A., and Gbenle, G. O. 2009.
Decolourization of Azo Dye by a Strain of Micrococcus Isolated from a
Refuse Dump Soil. Biotechnology. 8: 442–448.
Nakanishi, M., Yatome, C., Ishida, N., and Kitade, Y. 2001. Putative
ACP Phosphodiesterase Gene (acpD) Encodes an Azoreductase. Journal
of Biological Chemistry. 276: 46394–46399.
Kirby, N., Marchant, R. and McMullan, G. 2000. Decolourization of
Synthetic Textile Dyes by Phlebia tremellosa. FEMS Microbiology
Letters. 188: 93–96.
Ramalho, P. A., Cardoso, M. H., Paulo, A. C., and Ramalho, M. T. 2004.
Characterization of Azo Reduction Activity in a Novel Ascomycetes
Yeast Strain. Applied and Environmental Microbiology. 70: 2279–288.
McMullan, G., Meehan, C., Conneely, A., Kirby, N., Robinson, T.,
Nigam, P., Banat, I. M., Marchant, R. and Smyth, W. F. 2001. Microbial
Decolourisation and Degradation of Textile Dyes. Applied Microbiology
and Biotechnology. 56: 81–87.
Van Der Zee. 2002. Anaerobic Azo Dye Reduction. Doctorial Thesis,
Wageningen University
Puvaneswari, N., Muthukrishnan, J., and Gunasekaran, P. 2006. Toxicity
Assessment and Microbial Degradation of Azo Dyes. Indian Journal of
Experimental Biology. 44: 618–626.
Supaka, N., Juntongjin, K., Damronglerd, S., Delia, M. L., and
Strehaiano, P. 2004. Microbial Decolourisation of Reactive Azo Dyes in
a Sequential Anaerobic-Aerobic System. Chemical Engineering Journal.
: 169–176.
Maier, J., Kandelbauer, A., Erlacher, A., Cavaco-Paulo, A., and Gubitz,
G. M. 2004. A New Alkali-Thermostable Azoreductase from Bacillussp.
Strain SF. Applied and Environmental Microbiology. 70: 837–844.
Chong, C. S., Ibrahim, Z., Md Salleh, M., Abdul Rashid, N. A., Yahya,
A., and Wong, W. J. 2006. Decolourization of Azo Dye Direct Blue 15
Using Batch Culture of Klebsiella sp. Petroleum and Natural Resources
Process. Regional Conference of Post-graduate for Engineering and
Science, School of Post-graduate Studies, Universiti Teknologi Malaysia,
-27 July.
Russ, R., Rau, J., and Stolz, A. 2000. The Function of Cytoplasmic
Flavin Reductases In The Reduction of Azo Dyes By Bacteria. Applied
and Environmental Microbiology. 66: 1429–1434.
Cull, S. G., Holbrey, J. D., Vargas-Mora, V., Seddon, K. R., and Lye, G.
J. 2000. Room-temperature Ionic Liquids as Replacements For Organic
Solvents in Multiphase Bioprocess Operations. Biotechnology and
Bioengineering. 69: 227–233.
Wasserscheid, P. and Welton, T. 2002. Ionic Liquids in Synthesis. Berlin:
Wiley-VCH
Santafé, A. A., Doumèche, B., Blum, L. J., Girard-Egrot, A. P.,
Marquette, C. A. 2010. 1-Ethyl-3-methylimidazolium ethylsulfate/copper
Catalyst for the Enhancement of Glucose Chemiluminescent Detection:
Effects on Light Emission and Enzyme Activity. Analytical Chemistry.
: 2401–4.
Tavares, A. P. M., Rodriguez, O., and Macedo, E. A. 2008. Ionic Liquids
as Alternative Co-solvents for Laccase: Study of Enzyme Activity and
Stability. Biotechnology and Bioengineering. 101: 201–207
Yang, Z., Yue, Y. J., Huang, W. C., Zhuang, Z. M., Chen, Z. T., and
Xing, M. 2009. Importance of the Ionic Nature of Ionic Liquids in
Affecting Enzyme Performance. Journal of Biochemistry. 145: 355–364.
Noritomi, H. 2011. Protease-Catalyzed Synthetic Reactions in Ionic
Liquids, Ionic Liquids: Applications and Perspectives, Alexander
Kokorin (Ed.), ISBN: 978-953-307-248-7, InTech, Available from:
http://www.intechopen.com /articles/show/title/protease-catalyzedsynthetic-reactions-in-ionic-liquids.
Lowry, O. H., Rosenbrough, N. J., Farr, A. I., and Randall, R. J. 1951.
Protein Measurement with the Folin Phenol Reagent. Journal of Biology
and Chemistry. 193: 265–275.
Moosvi, S., Keharia, H., and Madamwar, D. 2005. Decolourization of
Textile Dye Reactive Violet 5 by a Newly Isolated Bacterial Consortium
RVM 11.1. World Journal of Microbiology and Biotechnology. 21: 667–
Gottlieb, A., Shaw, C., Smith, A., Wheatley, A., and Forsythe, S. 2001.
The Toxicity of Textile Reactive Azo Dyes after Hydrolysis and
Decolourisation. Journal of Biotechnology. 101: 49–56.
Wang, H., Zheng, X. W., Su, J. Q., Tian, Y., Xiong, X. J., and Zheng, T.
L. 2009. Biological Decolourisation of The Reactive Dyes Reactive
Black 5 by A Novel Isolated Bacterial Strain Enterobacter sp. EC3.
Journal of Hazardous Materials. 171: 654–659.
Seesuriyachan, P., Shinji, T., Ampin, K., Srikarnjana, K., Shuichiro, M.,
and Kenji, A. 2007. Metabolism of Azo Dyes By Lactobacillus Casei
TISTR 1500 and Effects of Various Factors on Decolourisation. Water
Research. 41: 985–992.
Choong, L. Y. 2004. Enzymatic Studies on Azoreductase from
Enterococcus Strain C1 that Decolourizes Azo Dyes. B. Sc. Thesis,
Universiti Teknologi Malaysia, Johor Bahru
Ooi, T., Shibata, T., Sato, R., Ohno, H., Kinoshita, S., and Thuoc, T.L. et
al. 2007. An Azoreductase, Aerobic NADH-Dependent Flavoprotein
Discovered From Bacillus sp. Functional Expression and Enzymatic
Characterization. Applied Microbiology and Biotechnology. 75: 377–386.
Panswad, T., and Luangdilok, W. 2000. Decolourisation of Reactive
Dyes with Different Molecular Structures Under Different Environmental
Conditions. Water Research. 34: 4177–4184.
Basso, A., Cantone, S., Linda, P., and Ebert, C. 2005. Stability and
Activity of Immobilsed Penicilin G Amidase In Ionic Liquids At
Controlled aw. Green Chemistry. 7: 671–676.
Yang, Z., Yue, Y., and Xing, M. 2008. Tyrosinase Activity in Ionic
Liquids. Biotechnology Letters. 30: 153–158.
Duplissa, L., Andreescu, S., Baltus, R., and Njagi, J. 2010. Interactions of
Room Temperature Ionic Liquids with Enzymes: Characterization and
Activity Studies. Chemical Engineering REU. 53: 432–436.
Downloads
Published
Issue
Section
License
Copyright of articles that appear in Jurnal Teknologi belongs exclusively to Penerbit Universiti Teknologi Malaysia (Penerbit UTM Press). This copyright covers the rights to reproduce the article, including reprints, electronic reproductions, or any other reproductions of similar nature.