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

Brazilian Archives of Biology and Technology

Ariane Leites Larentis²  Tito Lívio Moitinho Alves²  Orlando Bonifácio Martins¹ 

[1] ,Universidade Federal do Rio de Janeiro Centro de Tecnologia Programa de Engenharia Química - COPPE

关键词: Carbazole;    2'-aminobiphenyl-2;    3-diol dioxygenase;    HOADA;    heterologous gene expression;    petroleum denitrogenation;    biodegradation;   

DOI  :  10.1590/S1516-89132005000400016

来源: SciELO

【 摘 要 】

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 pDEST TM17 and salt induction for 4h.

【 授权许可】

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