Molecular cloning and partial characterization of the pathway for aniline degradation inPseudomonas sp. strain CIT1

The genes encoding aniline utilization inPseudomonas sp. strain CIT1 have been cloned inEscherichia coli and partially characterized. Molecular cloning of the genes was achieved by construction of a cosmid library, followed by mobilization of the library into mutants ofPs. sp. CIT1 impaired in a number of functions necessary for growth on aniline. A 42-kbSau3A fragment was found to encode the ability to utilize aniline and contained the catechol 2,3-dioxygenase (C230) gene. The regions encoding these activities were subcloned and further characterized. Plasmids containing the aniline oxidase gene encoded a 260-kDa protein complex, which was putatively shown to be composed of 72 kDa and possibly 36 kDa subunits. The fragment required for C230 activity encodes a 35 kDa protein, similar in size to C230 gene products previously characterized.