Formation Of Recombinant Strain Producing Catechol And Optimization Of Fermentation Conditions

Aniline is a widely used and poisonous compound, and its biodegrade has a huge industrial interest. In aerobic micoorganisms, aniline dioxygenase gene cluster plays an essential role in degradeing aniline by encoding the first enzyme in this metabolism, and catalyzes aniline to catechol. Catechol is an important compound and is used widely in industry. Producing catechol by microorganism has more advantages than by traditional methords.A gene library was constructed with the chromosomal DNA isolated from Acinetobacter sp. strain ANA5, this strain is able to utilize aniline as the sole carbon, nitrogen and energy source. A positive clone was selected by clone in situ hybridization and functional screening. A complete aniline dioxygenase gene cluster was obtained through subcloning and jointing. Sequence analysis showed that the gene cluster had six intact ORFs, ahead of which there was a promoter, and the whole sequence had high similarity with that of Acinetobacter sp. YAA at amino acid level. The gene cluster was cloned into a cosmid vector pLAFR6 which has no promoter to form...