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  Identification of a chlorobenzene reductive dehalogenase in dehalococcoides sp. strain CBDB1

Adrian, L., Rahnenführer, J., Gobom, J., & Hölscher, T. (2007). Identification of a chlorobenzene reductive dehalogenase in dehalococcoides sp. strain CBDB1. Applied and Environmental Microbiology, 73(23), 7717-7724. doi:10.1128/AEM.01649-07.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-8114-8 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-8115-6
Genre: Journal Article
Alternative Title : Appl Environ Microbiol

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 Creators:
Adrian, Lorenz, Author
Rahnenführer, Jan, Author
Gobom, Johan1, Author              
Hölscher, Tina, Author
Affiliations:
1Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, escidoc:1433550              

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 Abstract: A chlorobenzene reductive dehalogenase of the anaerobic dehalorespiring bacterium Dehalococcoides sp. strain CBDB1 was identified. Due to poor biomass yields, standard protein isolation procedures were not applicable. Therefore, cell extracts from cultures grown on trichlorobenzenes were separated by native polyacrylamide gel electrophoresis and analyzed directly for chlorobenzene reductive dehalogenase activity within gel fragments. Activity was found in a single band, even though electrophoretic separation was performed under aerobic conditions. Matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) and nano-liquid chromatography-MALDI MS analysis of silver-stained replicas of the active band on native polyacrylamide gels identified a protein product of the cbdbA84 gene, now called cbrA. The cbdbA84 gene is one of 32 reductive dehalogenase homologous genes present in the genome of strain CBDB1. The chlorobenzene reductive dehalogenase identified in our study represents a member of the family of corrinoid/iron-sulfur cluster-containing reductive dehalogenases. No orthologs of cbdbA84 were found in the completely sequenced genomes of Dehalococcoides sp. strains 195 and BAV1 nor among the genes amplified from Dehalococcoides sp. strain FL2 or mixed cultures containing Dehalococcoides. Another dehalogenase homologue (cbdbA80) was expressed in cultures that contained 1,2,4-trichlorobenzene, but its role is unclear. Other highly expressed proteins identified with our approach included the major subunit of a protein annotated as formate dehydrogenase, transporter subunits, and a putative S-layer protein.

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Language(s): eng - English
 Dates: 2007-12
 Publication Status: Published in print
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Title: Applied and Environmental Microbiology
  Alternative Title : Appl Environ Microbiol
Source Genre: Journal
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Pages: - Volume / Issue: 73 (23) Sequence Number: - Start / End Page: 7717 - 7724 Identifier: ISSN: 0099-2240