Identification of a novel dioxygenase involved in metabolism of o-xylene, toluene, and ethylbenzene by Rhodococcus sp. strain DK17

Kim, D; Chae, JC; Zylstra, GJ; Kim, YS; Kim, SK; Nam, MH; Kim, YM; Kim, E

HERO ID

1718856

Reference Type

Journal Article

Year

2004

Language

English

PMID

15574904

HERO ID 1718856
In Press No
Year 2004
Title Identification of a novel dioxygenase involved in metabolism of o-xylene, toluene, and ethylbenzene by Rhodococcus sp. strain DK17
Authors Kim, D; Chae, JC; Zylstra, GJ; Kim, YS; Kim, SK; Nam, MH; Kim, YM; Kim, E
Journal Applied and Environmental Microbiology
Volume 70
Issue 12
Page Numbers 7086-7092
Abstract Rhodococcus sp. strain DK17 is able to grow on o-xylene, benzene, toluene, and ethylbenzene. DK17 harbors at least two megaplasmids, and the genes encoding the initial steps in alkylbenzene metabolism are present on the 330-kb pDK2. The genes encoding alkylbenzene degradation were cloned in a cosmid clone and sequenced completely to reveal 35 open reading frames (ORFs). Among the ORFs, we identified two nearly exact copies (one base difference) of genes encoding large and small subunits of an iron sulfur protein terminal oxygenase that are 6 kb apart from each other. Immediately downstream of one copy of the dioxygenase genes (akbA1a and akbA2a) is a gene encoding a dioxygenase ferredoxin component (akbA3), and downstream of the other copy (akbA1b and akbA2b) are genes putatively encoding a meta-cleavage pathway. RT-PCR experiments show that the two copies of the dioxygenase genes are operonic with the downstream putative catabolic genes and that both operons are induced by o-xylene. When expressed in Escherichia coli, AkbA1a-AkbA2a-AkbA3 transformed o-xylene into 2,3- and 3,4-dimethylphenol. These were apparently derived from an unstable o-xylene cis-3,4-dihydrodiol, which readily dehydrates. This indicates a single point of attack of the dioxygenase on the aromatic ring. In contrast, attack of AkbA1a-AkbA2a-AkbA3 on ethylbenzene resulted in the formation of two different cis-dihydrodiols resulting from an oxidation at the 2,3 and the 3,4 positions on the aromatic ring, respectively.
Doi 10.1128/AEM.70.12.7086-7092.2004
Pmid 15574904
Wosid WOS:000225719300019
Url https://www.proquest.com/docview/17709873?accountid=171501&bdid=64564&_bd=Xshs44XCnO9GS3kmHqb4a2k%2FnPI%3D
Is Certified Translation No
Dupe Override No
Comments Source: Web of Science WOS:000225719300019
Is Public Yes
Language Text English