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Development of an enzymological (laccase-based) remediation product and technology (LACREMED)
Start date: Dec 31, 2011, End date: Dec 30, 2013 PROJECT  FINISHED 

Pesticides frequently pollute water and soils. Microbial degradation of pesticides like chlorinated anilinogenic and phenoligenic herbicides results in highly toxic aniline and phenol derivatives. These herbicides are frequently used mainly in grain cultures both in Hungary and in Serbia. Therefore, there is a substantial interest to tackle with challenges of soil and water quality, providing solid bases for sustainable farming and safe food production. This project will develop a new, innovative, enzyme (laccase)-based bioremediation product and application technology, enabling detoxification of soil and water with respect to the groups of phenol and aniline derivates. Developed product/technology takes into account cost efficiency of SMEs and farmers. Such product will be easily applied in water treating plants and on soils by end-users, enabling the preservation of the environmental quality in the regions known for its rich agricultural tradition. This 24-months cooperation will initiate long term joint research. Networking with SMEs in the region will enhance visibility of obtained results and the spread of the new bioremediation technology. Joint activity of researchers from Csongrád and South Bačka counties will target the soil and water quality, having ultimate goal to protect consumer health and preserving the environment. Hence, the population of the region living in this and other agricultural areas might benefit from the product, while competitiveness of the local agricultural producers could be enhanced in a long term on the EU market. Achievements: Laccase producer white-rot fungi from rotting wood logs were screened using different media andchromogenic substrates. Three different media were tested; best results were achieved with abasal medium. A chromogenic laccase substrate (guaiacol, 0,5 mg/ml) was added for theidentification of laccase activities. For the isolation of ascomyceteous laccase producers ABTS wasused in the media as chromogenic substrate. Altogether, 68 filamentous fungal strains able to produce laccase were isolated. Some of them were identified by microscopic investigations, others by sequencing. Laccase production of the selected isolates was measured in different liquid media. In some cases copper sulphate highly induced laccase production. Two media for the efficient isolation of laccaseproducing filamentous fungi were developed. Three methods for sensitive microtiter plate based laccase activity measuring methods were developed. The optimum pH values of the secreted laccases were measured from cell free tenfold diluted supernatants. In the experiments we used 5mM ABTS as substrate. On the basis of the experiments we could conclude that the optimal pH for all of the produced laccases is under pH 4.0.The effect of metal ions and the temperature to the activity of crude laccase enzyme extractswas tested. For the investigation of substrate spectra of laccases the cell free supernatants were used. The ferment broths were mixed with equal volume of the distinct xenobiotics. In some cases mediator (guaiacol) also was added to the reaction mixture. The mixtures were incubated at 25 °C overnight. The reactions were stopped by removing the laccase from the system, with membrane filtration.
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  • 85%   243 950,00
  • 2007 - 2013 Hungary - Serbia IPA CBC (HU-RS)
  • Project on KEEP Platform
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