Cloning and functional study of a novel aromatic-ring-hydroxylating dioxygenase gene.
The aromatic-ring-hydroxylating dioxygenase is a key enzyme that initiates the biodegradation of polycyclic aromatic hydrocarbons in bacteria. In the present study, a novel dioxygenase sequence was cloned from Terrabacter sp. FLO using a genome walking method. The dioxygenase was cloned into pET17 and actively expressed in E.coli BL21 (DE3) in co-expression with electron transfer chain proteins. The recombinant dioxygenase was found to transform phenanthrene, fluorene, pyrene and fluoranthene into the cis-dihydrodiol metabolites.
Zhou
Hong-wei
HW
Department of Enviromental health, School of Public Heath anf Tropical Medicine, Southern Medical University, Guangzhou, China. hzhou@fimmu.com
Zhou
Mei-juan
MJ
D000192:Actinomycetales / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005449:Fluorenes / Q000378:metabolism; D006900:Hydroxylation; D008969:Molecular Sequence Data; D010616:Phenanthrenes / Q000378:metabolism; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism*; D011721:Pyrenes / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D005449:Fluorenes; D010616:Phenanthrenes; D011084:Polycyclic Aromatic Hydrocarbons; D011721:Pyrenes; D011994:Recombinant Proteins; C007738:fluoranthene; C041509:fluorene; C031181:phenanthrene; C030984:pyrene; D049308:Dioxygenases