Das Projekt "The development of advanced bioremediation techniques for efficient treatment of industrial wastewaters" wird vom Umweltbundesamt gefördert und von Technische Universität Braunschweig, Institut für Mikrobiologie durchgeführt. The monitoring of the organic pollutants and the risk assessments in the Dnepropetrovsk region, Ukraine, will be studied on the territory of industrial enterprise 'Dneproshina' producing automotive tires as well on encircled area. The data obtained will result both in identification of the toxic compounds present in the studying material (industrial effluents and regions of the relevant industrial influence) and in ranking of these pollutants according to their danger from the ecological and population health points of view. Such data will also lead to estimation of the environmental impact of the pollutants discarded by industrial companies in the natural resources (Dnieper river and its basin, Black Sea). These analytical-ecological investigations will result in setting of the priorities for the development and application of the specific biotechnological methods for depolluting of the detected contaminants. In parallel, probes of the industrial effluents (both liquid and solid) will be analyzed on the presence of microorganisms capable to remediate of the found contaminants. Such microorganisms will be isolated, multiplied and then will be used for studies of optimal conditions for detoxification and mineralization of the detected pollutants. To perform the molecular assessment of in situ biodegradative capacity of the strains present in the effluents, the polymerase chain reaction (PCR) and reverse transcriptase-PCR (RT-PCR) techniques to measure the presence and expression of genes involved in the degradation of organic pollutants will be applied. The research plan foresees a sequencing and identification of novel genes encoding oxygenase enzymes involved in the degradation of aromatic pollutants as well as the application of forced evolution techniques to the generation of oxygenase enzymes with enhanced bioremediation capabilities. The selected genes will be integrated into the genomes of the parental or other appropriate recipient strains and genetically stable recombinants will be selected. These strains will be used for the analysis of depletion of xenobiotics under laboratory and simulated 'on-site' conditions. Their performance will be compared with that of the parental strains. To produce microorganism strains capable to biotransform the wide range of the pollutants two approaches will be used: microorganisms possessing polyfunctional ability to transform specific organic substances will be generated using both chromosomal integration and natural plasmid transfer; the microorganism consortia will be elaborated. The remediation efficiency of the obtained systems will be assessed. As the proposed research foresees the use of recombinant DNA technique, the special consideration will be given to biological safety studies. In this respect, the studies on possible gene transfer and plasmids'/integrated genes' stability are also planned. ...