The oxygenase component of the2-aminobenzenesulfonate dioxygenase system from Alcaligenes sp. strain O-1

Mampel, Jörg; Ruff, Jürgen; Junker, Frank; Cook, Alasdair M.

The oxygenase component of the2-aminobenzenesulfonate dioxygenase system from Alcaligenes sp. strain O-1

Dateien

The_oxygenase_component.pdfGröße: 344.16 KBDownloads: 361

Datum

1999

Autor:innen

Mampel, Jörg

Ruff, Jürgen

Junker, Frank

Cook, Alasdair M.

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Biologie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Microbiology. 1999, 145(11), pp. 3255-3264. ISSN 1350-0872. eISSN 1465-2080. Available under: doi: 10.1099/00221287-145-11-3255

Zusammenfassung

Growth of Alcaligenes sp. strain O-1 with 2-aminobenzenesulfonate (ABS; orthanilate) as sole source of carbon and energy requires expression of the soluble, multicomponent 2-aminobenzenesulfonate 2,3-dioxygenase system (deaminating) (ABSDOS) which is plasmid-encoded. ABSDOS was separated by anion-exchange chromatography to yield a flavin-dependent reductase component and an iron-dependent oxygenase component. The oxygenase component was purified to about 98% homogeneity and an {alpha}2ß2 subunit structure was deduced from the molecular masses of 134, 45 and 16 kDa for the native complex, and the {alpha} and ß subunits, respectively. Analysis of the amount of acid labile sulfur and total iron, and the UV spectrum of the purified oxygenase component indicated one [2Fe 2S] Rieske centre per {alpha} subunit. The inhibition of activity by the iron-specific chelator o-phenanthroline indicated the presence of an additional iron-binding site. Recovery of active protein required strictly anoxic conditions during all purification steps. The FAD-containing reductase could not be purified. ABSDOS oxygenated nine sulfonated compounds; no oxygen uptake was detected with carboxylated aromatic compounds or with aliphatic sulfonated compounds. Km values of 29, 18 and 108 µM and Vmax values of 140, 110 and 72 pkat for ABS, benzenesulfonate and 4-toluenesulfonate, respectively, were observed. The N-terminal amino acid sequences of the {alpha}- and ß-subunits of the oxygenase component allowed PCR primers to be deduced and the DNA sequence of the {alpha}-subunit was thereafter determined. Both redox centres were detected in the deduced amino acid sequence. Sequence data and biochemical properties of the enzyme system indicate a novel member of the class IB ring-hydroxylating dioxygenases.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Schlagwörter

Alcaligenes sp. strain O-1, characterization of oxygenase, 2-aminobenzenesulfonate dioxygenase system, oxygenase sequence

Zitieren

ISO 690

MAMPEL, Jörg, Jürgen RUFF, Frank JUNKER, Alasdair M. COOK, 1999. The oxygenase component of the2-aminobenzenesulfonate dioxygenase system from Alcaligenes sp. strain O-1. In: Microbiology. 1999, 145(11), pp. 3255-3264. ISSN 1350-0872. eISSN 1465-2080. Available under: doi: 10.1099/00221287-145-11-3255

BibTex

@article{Mampel1999oxyge-7876,
  year={1999},
  doi={10.1099/00221287-145-11-3255},
  title={The oxygenase component of the2-aminobenzenesulfonate dioxygenase system from Alcaligenes sp. strain O-1},
  number={11},
  volume={145},
  issn={1350-0872},
  journal={Microbiology},
  pages={3255--3264},
  author={Mampel, Jörg and Ruff, Jürgen and Junker, Frank and Cook, Alasdair M.}
}

RDF

<rdf:RDF
    xmlns:dcterms="http://purl.org/dc/terms/"
    xmlns:dc="http://purl.org/dc/elements/1.1/"
    xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#"
    xmlns:bibo="http://purl.org/ontology/bibo/"
    xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#"
    xmlns:foaf="http://xmlns.com/foaf/0.1/"
    xmlns:void="http://rdfs.org/ns/void#"
    xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > 
  <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/7876">
    <dc:creator>Cook, Alasdair M.</dc:creator>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dcterms:abstract xml:lang="eng">Growth of Alcaligenes sp. strain O-1 with 2-aminobenzenesulfonate (ABS; orthanilate) as sole source of carbon and energy requires expression of the soluble, multicomponent 2-aminobenzenesulfonate 2,3-dioxygenase system (deaminating) (ABSDOS) which is plasmid-encoded. ABSDOS was separated by anion-exchange chromatography to yield a flavin-dependent reductase component and an iron-dependent oxygenase component. The oxygenase component was purified to about 98% homogeneity and an {alpha}2ß2 subunit structure was deduced from the molecular masses of 134, 45 and 16 kDa for the native complex, and the {alpha} and ß subunits, respectively. Analysis of the amount of acid labile sulfur and total iron, and the UV spectrum of the purified oxygenase component indicated one [2Fe 2S] Rieske centre per {alpha} subunit. The inhibition of activity by the iron-specific chelator o-phenanthroline indicated the presence of an additional iron-binding site. Recovery of active protein required strictly anoxic conditions during all purification steps. The FAD-containing reductase could not be purified. ABSDOS oxygenated nine sulfonated compounds; no oxygen uptake was detected with carboxylated aromatic compounds or with aliphatic sulfonated compounds. Km values of 29, 18 and 108 µM and Vmax values of 140, 110 and 72 pkat for ABS, benzenesulfonate and 4-toluenesulfonate, respectively, were observed. The N-terminal amino acid sequences of the {alpha}- and ß-subunits of the oxygenase component allowed PCR primers to be deduced and the DNA sequence of the {alpha}-subunit was thereafter determined. Both redox centres were detected in the deduced amino acid sequence. Sequence data and biochemical properties of the enzyme system indicate a novel member of the class IB ring-hydroxylating dioxygenases.</dcterms:abstract>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:creator>Mampel, Jörg</dc:creator>
    <dc:contributor>Mampel, Jörg</dc:contributor>
    <dc:contributor>Cook, Alasdair M.</dc:contributor>
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by-nc-nd/2.0/"/>
    <dc:creator>Junker, Frank</dc:creator>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dcterms:bibliographicCitation>First publ. in: Microbiology 145 (1999), 11, pp. 3255-3264</dcterms:bibliographicCitation>
    <dc:creator>Ruff, Jürgen</dc:creator>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:38:12Z</dc:date>
    <dc:contributor>Junker, Frank</dc:contributor>
    <dc:contributor>Ruff, Jürgen</dc:contributor>
    <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/7876"/>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:38:12Z</dcterms:available>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/7876/1/The_oxygenase_component.pdf"/>
    <dcterms:issued>1999</dcterms:issued>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/7876/1/The_oxygenase_component.pdf"/>
    <dcterms:title>The oxygenase component of the2-aminobenzenesulfonate dioxygenase system from Alcaligenes sp. strain O-1</dcterms:title>
    <dc:language>eng</dc:language>
    <dc:format>application/pdf</dc:format>
    <dc:rights>Attribution-NonCommercial-NoDerivs 2.0 Generic</dc:rights>
  </rdf:Description>
</rdf:RDF>

Universitätsbibliographie

Nein