Decolouration of azo dyes by Phanerochaete chrysosporium immobilised into alginate beads

Because of high discharged volumes and effluent composition, wastewater from the textile industry can be considered as the most polluting amongst all industrial sectors, thus greatly requiring appropriate treatment technologies. Although some abiotic methods for the reduction of several dyes exist, these require highly expensive catalysts and reagents. Biotechnological approaches were proven to be potentially effective in the treatment of this pollution source in an eco-efficient manner. The white-rot fungi are, so far, the most efficient microorganisms in degrading synthetic dyes. This white-rot fungi's property is due to the production of extracellular lignin-modifying enzymes, which are able to degrade a wide range of xenobiotic compounds because of their low substrate specificity. In this paper, we studied the ability of the white-rot fungus Phanerochaete chrysosporium immobilised into Ca-alginate beads to decolourise different recalcitrant azo dyes such as Direct Violet 51 (DV), Reactive Black 5 (RB), Ponceau Xylidine (PX) and Bismark Brown R (BB) in successive batch cultures. To the best of our knowledge, this is the first study on the immobilisation of P. chrysosporium into Ca-alginate beads for its application in dye decolouration.

P. chrysosporium was immobilised into Ca-alginate beads using a method of gel recoating to minimise cellular leaking. The immobilised fungus was transferred to 250-ml Erlenmeyer flasks containing 50 ml of growth medium and incubated on an orbital shaker at 150 rpm and 30 degrees C for 7 days. The ratio of beads/medium used was 10% (w/v). The dyes were added into the culture flasks when MnP production started (50 U l(-1)), which corresponded with the seventh cultivation day. MnP activity and dye decolouration were measured spectrophotometrically.

The dyes DV, RB and PX were almost totally decolourised at the end of each batch during the course of three successive batches. However, the dye BB was more resistant to decolouration and it was not completely decolourised (86.7% in 144 h). Further, the beads were kept in sterilised calcium chloride (2 g l(-1)) for 3 weeks at 4 degrees C. After these three storage weeks, the immobilised P. chrysosporium was again efficiently reused for azo dye decolouration during two successive batches, decolouration being more effective even for BB. Also, the in vitro decolouration of the aforementioned azo dyes by crude MnP from P. chrysosporium was performed. The decolouration levels obtained were lower than those attained with the whole cultures especially for RB and BB dyes, in spite of the fact that dye concentrations used were considerable lower.

The good performance of the immobilisation system was likely due to the gel re-coating method utilised to prepare the alginate beads which not only maintained the beads integrity but also avoided cellular leaking. The lower decolouration percentages obtained by the enzyme indicates that the mycelial biomass may supply other intracellular or mycelial-bound enzymes, or other compounds that favour dye decolouration.

Immobilised P. chrysosporium efficiently decolourised different types of azo dyes. In this decolouration process, the MnP secreted by the fungus played the main role whilst adsorption was found to be negligible except for the dye BB.

Efforts should be made to scale up and apply fungal decolouration techniques to real industrial dye-containing wastewater. Further, detailed characterisation of the intermediates and metabolites produced during biodegradation must be done to ensure the safety of the decolourised wastewater.