5. The final goal will consist in the application of bioremediation directly in the endangered regions. On the basis of the previous studies and results those strains, either natural or engineered, which have been recognized able to degrade hydrocarbons will be used in situ and their effectiveness will be tested. All the factors limiting degradation rates in biodegradation be adequately identified and addressed. The effectiveness of bioremediation should be tested not only for the decrease of the hydrocarbon concentration, but also for the decrease of toxicity to ensure that additional toxicity of metabolites is insignificant.
The aim of the following proposal is the application of natural resources for recovering the Caspian area from hydrocarbons's pollution.
Specific goals of the proposal are identified in the following points :
1. Monitoring of the composition of oils which are produced in the Caspian region, the identification of areas of vulnerability for spills, that in principle should correspond to the regions of oil-fields and pipelines and of the type of hydrocarbons responsible of the pollution of both the sea and the coast.
2. Isolation of organisms able to grow and to use as carbon source hydrocarbon substrates. Some hydrocarbon-degrading bacteria have been recognized to belong to the classes of Achromobacter, Acinetobacter, Alcaligens, Arthrobacter, Bacillus, Flavobacterium, Nocordia and Pseudomonas spp. Of course, some of them are expected to be present also in the Caspian environment. Furthermore, new types of organisms, with similar or more efficient degrading characteristics, could be identified.
3. Monitoring in situ of biodegradation activity is important in order to determine whether bioaugmentation or nutrient addition is worthwhile and to determine the effectiveness of any such biological treatment. Recently, methods have been developed for monitoring in situ catabolic gene expression using direct isolation of mRNA from contaminated soil. A very sensitive technique of reverse transcriptase-polymerase chain reaction (RT-PCR) (Becker-André, M. and Hahibrock, K. (1989) Nucl. Acid. Res. 17:9437-9446) will be applied to the quantification of specific mRNAs from environmental samples.
4. The next goal must concern the understanding of the molecular aspects of biodegradation and to detect the degradative pathways of the hydrocarbons which are present in that environment. This knowledge is particularly relevant for a real application of bioremediative technique and for possible improvements of degradative efficiency of bacterial strains through genetic manipulations. A large number of hydrocarbons are degraded by bacterial communities which contain multienzymatic systems. These enzymes will be isolated and their active site characterized using a variety of spectroscopic techniques like NMR, EPR, CD, MCD, electronic absorption. Crystallization trials will be performed in order to obtain crystal for X-ray structure determination. The specificity of these enzymes will also be tested: the interaction with different substrates will be monitored through kinetic and spectroscopic measurements.
Funding SchemeCSC - Cost-sharing contracts
SE1 9NN London