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PMID: 9925607 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The chlorocatechol-catabolic transposon Tn5707 of Alcaligenes eutrophus NH9, carrying a gene cluster highly homologous to that in the 1,2,4-trichlorobenzene-degrading bacterium Pseudomonas sp. strain P51, confers the ability to grow on 3-chlorobenzoate.

Applied and environmental microbiology ·Vol. 65 ·No. 2 ·1999-02-00 ·Pages 724-31

Ogawa N, Miyashita K

Abstract

Alcaligenes eutrophus (Ralstonia eutropha) NH9, isolated in Japan, utilizes 3-chlorobenzoate as its sole source of carbon and energy. Sequencing of the relevant region of plasmid pENH91 from strain NH9 revealed that the genes for the catabolic enzymes were homologous to the genes of the modified ortho-cleavage pathway. The genes from strain NH9 (cbnR-ABCD) showed the highest homology (89 to 100% identity at the nucleotide level) to the tcbR-CDEF genes on plasmid pP51 of the 1,2,4-trichlorobenzene-degrading bacterium Pseudomonas sp. strain P51, which was isolated in The Netherlands. The structure of the operon, including the lengths of open reading frames and intervening sequences, was completely conserved between the cbn and tcb genes. Most nucleotide substitutions were localized within and proximal to the cbnB (tcbD) gene. The difference in the chloroaromatics that the two strains could use as growth substrates seemed to be due to differences in enzymes that convert substrates to chlorocatechols. The restriction map of plasmid pENH91 was clearly different from that of pP51 except in the regions that contained the cbnR-ABCD and tcbR-CDEF genes, respectively, suggesting that the chlorocatechol gene clusters might have been transferred as units. Two homologous sequences, present as direct repeats in both flanking regions of the cbnR-ABCD genes on pENH91, were found to be identical insertion sequences (ISs), designated IS1600, which formed a composite transposon designated Tn5707. Although the tcbR-CDEF genes were not associated with similar ISs, a DNA fragment homologous to IS1600 was cloned from the chromosome of strain P51. The sequence of the fragment suggested that it might be a remnant of an IS. The two sequences, together with IS1326 and nmoT, formed a distinct cluster on a phylogenetic tree of the IS21 family. The diversity of the sources of these IS or IS-like elements suggests the prevalence of ISs of this type.

MeSH Terms
Amino Acid Sequence Base Sequence Biodegradation, Environmental Catechols/metabolism Chlorobenzenes/metabolism Chlorobenzoates/metabolism Cloning, Molecular Cupriavidus necator/enzymology,genetics DNA Transposable Elements Genes, Bacterial Molecular Sequence Data Multigene Family Open Reading Frames Phylogeny Pseudomonas/enzymology,genetics Restriction Mapping Sequence Analysis, DNA Sequence Homology, Amino Acid Sequence Homology, Nucleic Acid
Chemicals
Catechols Chlorobenzenes Chlorobenzoates DNA Transposable Elements 3-chlorobenzoic acid 1,2,4-trichlorobenzene
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ogawa N
National Institute of Agro-Environmental Sciences, 3-1-1 Kan-nondai, Tsukuba, Ibaraki 305-8604, Japan. naotow@niaes.affrc.go.jp
Miyashita K
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.65.2.724-731.1999
Email
naotow@niaes.affrc.go.jp
Published
1999-02-00
Pages
724-31
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC91086
Subset
IM
Databases
GENBANK
AB019032, AB019033, D64144
External Links
PubMed source DOI: 10.1128/AEM.65.2.724-731.1999 PMC Full Text View journal details
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