Kinetics of biological decolorisation of anthraquinone based Reactive Blue 19 using an isolated strain of Enterobacter sp.F NCIM 5545
Introduction
The textile industry consumes large quantities of water and hence produces large volumes of wastewater from different steps in the dyeing and finishing processes. Wastewater from printing and dyeing units is often rich in color, containing residues of dyes. Textile wastewater pollutants generally contain dyes that increase its BOD, COD, solid contents, color and toxicity. This water must be treated before discharge in environment. For the treatment, removal color (i.e., breakage of chromophore) is necessary. The most important chromophores are azo (NN), carbonyl (CO), nitro (NO) and quinoid groups. Azo and anthraquinone are the most important chromophoric groups from which textile dyes are synthesized (He et al., 2008). It is believed that anthraquinone-based dyes are more resistant to biodegradation due to their fused aromatic structures compared to azo-based ones (He et al., 2008, Fanchiang and Tseng, 2009). Extensive studies have been carried out in the decolorization of azo dyes but there are very few studies reported for an anthraquinone dye.
For decolorisation or degradation, treatment methods generally involve various oxidation methods (cavitation (Gogate and Mishra, 2010, Pandit et al., 2011, Pandit et al., 2013), photocatalytic oxidation, ozone, H2O2, Fenton’s chemistry (Abdullah et al., 2007)), physical methods (flocculation, adsorption (Kumar and Ahmad, 2011)) and biological methods (fungi (Mohammad et al., 2009), algae (Khataee et al., 2011) and bacteria (Ponraj et al., 2011)). Physical/chemical methods are dealing with dye wastewater treatment but these methods have some drawbacks, such as inability to completely remove the recalcitrant dyes and generation of significant amount of sludge that may cause secondary pollution problems; substantially increases the cost of these treatment methods (Pearce et al., 2003). Compared to physical and chemical methods, biological processes are time consuming but result into complete mineralization of dye (Rai et al., 2005). Within biological processes, compared to algae and fungus, bacterial decolorisation requires less hydraulic retention time (Alalewi and Jiang, 2012).
Reactive Blue-19 (RB-19), also known as Remazol brilliant blue is an anthraquinone based vinyl sulphone dye, which is very resistant to chemical oxidation due to its anthraquinone structure being stabilized by resonance (Fanchiang and Tseng, 2009). RB-19 has relatively low fixation ability (75–80%) due to the competition between the formation of vinyl sulphone and the hydrolysis reactions, i.e., formation of 2-hydroxysulphone, which does not attach to the fiber as a result that dye remains stable into the wastewater for a long period of time, i.e., half life of RB-19 is 46 years at pH 7 and at a temperature of 25 °C that renders transport resistance to the oxygen transfer to aquatic life (Memon and Memon, 2012). Therefore, the strategy to remove the color and to decrease its impact on the wastewater of textile environment is of significant importance.
Thus, the bacterial decolorisation of anthraquinone dye (C.I. Reactive Blue 19) by a newly isolated strain of Enterobacter sp.F have been reported in the present work. Kinetics of biological decolorisation of anthraquinone based Reactive Blue 19 have also been evaluated.
Section snippets
Dyes and chemicals
Glucose, nutrient broth and agar were obtained from Hi-Media Pvt. Ltd. (India). The textile dyes: Reactive Blue 19 was kindly donated by Colorband Dyestuff (P) Ltd., INDIA.
Organism and culture condition
The organism used in this study was isolated mixed culture from anaerobic digester for biogas production from Institute of Chemical Technology, Mumbai, India. It was then maintained on nutrient agar slants at 4 °C. The pure culture was grown in 250 ml Erlenmeyer flask containing 100 ml nutrient broth containing beef extract 1 g/L
Statistical analysis
All the experiment and analysis were conducted in triplicate and results presented here are the mean of triplicate ± standard deviations (SD). For the comparison of two levels of a single factor, Statistical analysis was done using student “t” test by Graph pad software. For the comparison of three or more than three levels of a single factor, data were analyzed by one-way analysis of variance (ANOVA) by using Tukey–Kramer multiple comparisons test (GraphPad Prism version 6.0 for Windows, //www.graphpad.com/demos/download.cfm?demo
Isolation and identification of decolorizing bacteria
A bacterial strain, which had high decolorization ability against Reactive Blue 19, was isolated. Significant decolorization efficiency (90%) was achieved after incubation for 108 h. The colony of bacterial isolate F was filamentous and yellow–white. The strain F was observed to be a Gram-negative.
To analyze the phylogenetic position, the 16 S rDNA sequence of the strain F was determined. Fig. 1 showed the phylogenetic relationship between the strain F and other related microorganisms found in
Decolorisation mechanism of Enterobacter sp.F against Reactive Blue 19
Decolorisation of dye solution by bacteria could be due to adsorption to (a) microbial cell or (b) biochemical reaction or both leading to biodegradation. In adsorption, examination of the absorption spectrum would reveal that all peaks decreased approximately in the same proportion to each other. If dye removal is attributed to biochemical reaction, either the major visible light absorbance peak should have completely disappeared or a new peak would have appeared (Ayed et al., 2010).
In the
Biodegradation analysis
The degradation was confirmed from FTIR analysis. Comparison of FTIR spectrum of control dye with extracted metabolites after complete decolorisation, clearly indicated the biodegradation of the parent dye compound by Enterobacter sp.F.
The FTIR spectra revealed that the ring structures of RB-19 were definitely degraded because the peaks of the wavenumber between 670 and 870 cm−1, which denote aromatic rings, which was also disappeared (Fanchiang and Tseng, 2009). Moreover, the cleavage of
Conclusions
The optimal conditions for the decolorizing activity of Enterobacter sp.F were anaerobic conditions with 6 g/L glucose supplementation, 40 as glucose to microbe weight ratio, pH of 10, and temperature of 37 °C. Enterobacter sp.F showed decolorizing activity for RB 19 dye degradation through bio-chemical mechanism, and it could tolerate up to 400 mg/L of RB 19 without experiencing significant toxicity.
Because of degradative and decolorizing activity against anthraquinone reactive dye commonly used
Acknowledgements
Chandrakant Holkar gratefully acknowledges the Department of Bioprocess Technology (DBT), India for the Research Fellowship to carry out the work. The authors would like to thank Dr. D.V. Gokhale and Dr. Syed Dastager Scientist in NCIM Resource Centre, CSIR-NCL, Pune for the help in an identification of isolated strain.
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