Fungal laccases are enzymes that have been studied because of their ability to decolorize and detoxify effluents; they are also used in paper bleaching, synthesis of polymers, bioremediation, etc. In this work we were able to express a laccase from Trametes (Pycnoporus) sanguineus in the filamentous fungus Trichoderma atroviride. For this purpose, a transformation vector was designed to integrate the gene of interest in an intergenic locus near the blu17 terminator region. Although monosporic selection was still necessary, stable integration at the desired locus was achieved. The native signal peptide from T. sanguineus laccase was successful to secrete the recombinant protein into the culture medium. The purified, heterologously expressed laccase maintained similar properties to those observed in the native enzyme (Km and kcat and kcat/km values for ABTS, thermostability, substrate range, pH optimum, etc). To determine the bioremediation potential of this modified strain, the laccase-overexpressing Trichoderma strain was used to remove xenobiotic compounds. Phenolic compounds present in industrial wastewater and bisphenol A (an endocrine disruptor) from the culture medium were more efficiently removed by this modified strain than with the wild type. In addition, the heterologously expressed laccase was able to decolorize different dyes as well as remove benzo[α]pyrene and phenanthrene in vitro, showing its potential for xenobiotic compound degradation.
In Canada, the presence of pharmaceuticals and pesticides in municipal drinking water has been examined primarily in larger urban centres which draw their supplies from surface water. However, few studies have examined this issue in smaller and rural communities, which represent nearly one-third of the Canadian population and which draw their drinking water mainly from groundwater. This study presents a regional-scale assessment of the presence of these contaminants in the drinking waters of 17 smaller rural communities, compared with two larger urban communities, in south-central Quebec. From a total of 70 chemicals examined, 15 compounds (nine pharmaceuticals and six pesticides) were detected. The three most frequently detected contaminants were caffeine, atrazine and naproxen, respectively, in 29%, 24% and 21% of the samples. Detections reported here for the first time in Quebec drinking water include the known human carcinogen cyclophosphamide and the fungicide thiabendazole. Maximum concentrations of pharmaceuticals ranged from 30 to 1,848 ng L−1 and of pesticides from 21 to 856 ng L−1. This study provides direct evidence that drinking water in smaller, rural communities of Quebec, Canada, whether sourced from groundwater or surface water, can contain measurable levels of pharmaceuticals and pesticides, indicative of their susceptibility to source contamination.
This article has been made Open Access thanks to the kind support of CAWQ/ACQE (https://www.cawq.ca).
The potential of microorganisms for the treatment of municipal biosolids is continuously growing. The present studies evaluated the potency of Trametes hirsuta for the reduction in biosolid mass, production of extracellular enzymes, and removal of pharmaceutical compounds (PhACs) in biosolid slurry in the presence and absence of spiked PhACs [5 non-steroidal anti-inflammatories (NSAIs) and 2 psychoactive compounds (PACs)]. Toxicity after 35 days of fungal treatment was also assessed. Results showed that the growth of T. hirsuta is limited above 25% and wholly inhibited above 50% of biosolids in the slurry. At 12% of biosolid concentration, biosolid mass was reduced by 90%, NSAIs were entirely removed, but PACs' removal was only ~20%. Increasing biosolid content to 25% did not markedly affect biosolid reduction but significantly enhanced the removal of PACs (>50%). Results also showed that both PhACs and biosolids induced the production of oxidative enzymes. In 12% biosolids in the slurry, the oxidative potential measured by the ABTS assay (OABTS) reached 5,000 mM of OABTS in the presence of PhACs, and 2,500 mM of OABTS without PhACs, as compared to 1,200 mM of OABTS in control culture. Finally, we report that white rot fungi (WRF) treatment significantly decreased the toxicity of the biosolids.
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