Genetics of alkane degradation in pseudomonas oleovorans. Cel engineering for biotechnological applications.

The catabolic plasmid OCT extends the substrate range of P. putida from alkanols to n-alkanes and n-alkenes with a chain length between C6 and C12. The alk regulon, that is localized on this plasmid, consists of two distinct regions, the alkBAC operon and the AlkR locus which positively controls the expression of the alkBAC operon. The alkBAC operon encodes for two enzymatic functions: alkane hydroxylation and oxidation of the resulting alkanol. The oxidation of aliphatic compounds by P. oleovorans (P. putida carrying the OCT plasmid) is of interest due to the specific characteristics of this organism which grows in the presence of a bulk apolar phase and due to the potential utility of terminal oxidation in the production of alkanols, aldehydes, dicarboxylic acids, chiral epoxides and polyesters.
The research described in this thesis was aimed at a detailed genetic characterization of the genes responsible for alkane oxidation. Since the large 400-500 kb OCT plasmid which carries the alk genes cannot be handled for in vitro characterization and manipulation, a gene library was established with total P. oleovorans DNA. ...

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Summary and general conclusions

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