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Volume 145, Issue 11

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

The GenBank accession number for the sequence reported in this paper is AF109074.

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Abstract

Growth of 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 αβ subunit structure was deduced from the molecular masses of 134, 45 and 16 kDa for the native complex, and the α and β 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 α subunit. The inhibition of activity by the iron-specific chelator -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. values of 29, 18 and 108 μM and values of 140, 110 and 72 pkat for ABS, benzenesulfonate and 4-toluenesulfonate, respectively, were observed. The N-terminal amino acid sequences of the α- and β-subunits of the oxygenase component allowed PCR primers to be deduced and the DNA sequence of the α-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.

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Keyword(s): 2-aminobenzenesulfonate dioxygenase system , 3SCDO, 3-sulfocatechol dioxygenase , ABS, 2-aminobenzenesulfonate , ABSDOS, 2-ABS dioxygenase system , Alcaligenes sp. strain O-1 , BS, benzenesulfonate , characterization of oxygenase , oxygenase sequence , oxygenation and deamination and TS, 4-toluenesulfonate
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1999-11-01
2024-04-18
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The oxygenase component of the2-aminobenzenesulfonate dioxygenase system from Alcaligenes sp. strain O-1The GenBank accession number for the sequence reported in this paper is AF109074.
Microbiology 145, 3255 (1999); https://doi.org/10.1099/00221287-145-11-3255
/content/journal/micro/10.1099/00221287-145-11-3255
/content/journal/micro/10.1099/00221287-145-11-3255
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