Cloning and expression of meta-cleavage enzyme (CarB) of carbazole degradation pathway from Pseudomonas stutzeri

in this work, the 1082bp pcr product corresponding to carbabb genes that encode the heterotetrameric enzyme 2'-aminobiphenyl-2,3-diol 1,2-dioxygenase (carb), involved in the pseudomonas stutzeri atcc 31258 carbazole degradation pathway, was cloned using the site-specific recombination system. recombinant clones were confirmed by pcr, restriction enzyme digestion and sequencing. carb dioxygenase was expressed in high levels and in active form in escherichia coli bl21-si using the his-tagged expression vector pdesttm17 and salt induction for 4h.