Cloning and expression of thermophilic catechol 1,2-dioxygenase gene (catA) from Streptomyces setonii

An, Hyungjun

doi:10.1016/s0378-1097(00)00538-3

Cited by 7 publications

(17 citation statements)

References 14 publications

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“…Streptomyces setonii (ATCC 39116), originally isolated from vanillate‐enriched Idaho soil, degrades various single aromatic compounds including phenol and benzoate through a catechol intermediate via an ortho ‐cleavage pathway using C12O [17,18]. Previously, we constructed an S. setonii DNA library and isolated a 6.3‐kb Pst I fragment, within which a 1.4‐kb fragment containing the full‐length C12O gene, catA , was sequenced [19]. Nucleotide sequencing analysis of the 1.4‐kb DNA fragment revealed a 0.84‐kb open reading frame (ORF), which showed a strong overall amino acid similarity to the known high‐G+C Gram‐positive (but significantly less to the Gram‐negative) bacterial mesophilic C12Os [19].…”

Section: Introductionmentioning

confidence: 99%

“…Previously, we constructed an S. setonii DNA library and isolated a 6.3‐kb Pst I fragment, within which a 1.4‐kb fragment containing the full‐length C12O gene, catA , was sequenced [19]. Nucleotide sequencing analysis of the 1.4‐kb DNA fragment revealed a 0.84‐kb open reading frame (ORF), which showed a strong overall amino acid similarity to the known high‐G+C Gram‐positive (but significantly less to the Gram‐negative) bacterial mesophilic C12Os [19]. The heterologous expression of the cloned 1.4‐kb DNA fragment in Escherichia coli demonstrated that this C12O possessed a thermophilic activity within a broad temperature range (up to 65°C) and showed a higher activity against 3‐methylcatechol than catechol or 4‐methylcatechol, but no activity against protocatechuate [19].…”

Section: Introductionmentioning

confidence: 99%

“…Nucleotide sequencing analysis of the 1.4‐kb DNA fragment revealed a 0.84‐kb open reading frame (ORF), which showed a strong overall amino acid similarity to the known high‐G+C Gram‐positive (but significantly less to the Gram‐negative) bacterial mesophilic C12Os [19]. The heterologous expression of the cloned 1.4‐kb DNA fragment in Escherichia coli demonstrated that this C12O possessed a thermophilic activity within a broad temperature range (up to 65°C) and showed a higher activity against 3‐methylcatechol than catechol or 4‐methylcatechol, but no activity against protocatechuate [19]. In this report, the flanking regions of the S. setonii C12O gene were further investigated revealing for the first time among streptomycetes, the presence of divergently transcribed operons: for benzoate and catechol catabolic gene clusters.…”

Section: Introductionmentioning

confidence: 99%

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An inducibleStreptomycesgene cluster involved in aromatic compound metabolism

Park

Kim

2003

FEMS Microbiology Letters

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Streptomyces setonii (ATCC 39116) is a thermophilic soil actinomycete capable of degrading single aromatic compounds including phenol and benzoate via the ortho-cleavage pathway. Previously, a 6.3-kb S. setonii DNA fragment containing a thermophilic catechol 1,2-dioxygenase (C12O) gene was isolated and functionally overexpressed in Escherichia coli (An et al., FEMS Microbiol. Lett. 195 (2001) 17-22). Here the 6.3-kb S. setonii DNA fragment was shown to be organized into two putative divergently transcribed gene clusters with six complete and one incomplete open reading frames (ORFs). The first cluster with three ORFs showed homologies to previously known benA, benB, and benC, implying it is a part of the benzoate catabolic operon. The second cluster revealed an ortho-cleavage catechol catabolic operon with three translationally coupled ORFs (in order): catR, a putative LysR-type regulatory gene; catB, a muconate cycloisomerase gene; catA, a C12O gene. Each of these individually cloned ORFs was expressed in E. coli and identified as a distinct protein. The expression of the cloned S. setonii catechol operon was induced in Streptomyces lividans by specific single aromatic compounds including catechol, phenol, and 4-chlorophenol. A similar induction pattern was also observed using a luciferase gene-fused reporter system.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An inducibleStreptomycesgene cluster involved in aromatic compound metabolism

Park

Kim

2003

FEMS Microbiology Letters

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“…The gentisate pathway appears to be less common than the catechol pathway. The benzene ring of catechol is cleaved by a 1,2-dioxygenase or 2,3-dioxygenase (1,40), whereas gentisate is cleaved by gentisate 1,2-dioxygenase (GDO) (8,18,21,62,67). Investigations of salicylate degradation in gram-positive bacteria, including Streptomyces spp., have revealed the presence of both pathways (26).…”

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confidence: 99%

Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46

Ishiyama

Vujaklija

Davies

2004

Appl Environ Microbiol

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A novel salicylate-degrading Streptomyces sp., strain WA46, was identified by UV fluorescence on solid minimal medium containing salicylate; trace amounts of gentisate were detected by high-pressure liquid chromatography when strain WA46 was grown with salicylate. PCR amplification of WA46 DNA with degenerate primers for gentisate 1,2-dioxygenase (GDO) genes produced an amplicon of the expected size. Sequential PCR with nested GDO primers was then used to identify a salicylate degradation gene cluster in a plasmid library of WA46 chromosomal DNA. The nucleotide sequence of a 13.5-kb insert in recombinant plasmid pWD1 (which was sufficient for the complete degradation of salicylate) showed that nine putative open reading frames (ORFs) (sdgABCDEFGHR) were involved. Plasmid pWD1 derivatives disrupted in each putative gene were transformed into Streptomyces lividans TK64. Disruption of either sdgA or sdgC blocked salicylate degradation; constructs lacking sdgD accumulated gentisate. Cell extracts from Escherichia coli DH5␣ transformants harboring pUC19 that expressed each of the sdg ORFs showed that conversions of salicylate to salicylylcoenzyme A (CoA) and salicylyl-CoA to gentisyl-CoA required SdgA and SdgC, respectively. SdgA required CoA and ATP as cofactors, while NADH was required for SdgC activity; SdgC was identified as salicylyl-CoA 5-hydroxylase. Gentisyl-CoA underwent spontaneous cleavage to gentisate and CoA. SdgA behaved as a salicylyl-CoA ligase despite showing amino acid sequence similarity to an AMP-ligase. SdgD was identified as a GDO. These results suggest that Streptomyces sp. strain WA46 degrades salicylate by a novel pathway via a CoA derivative. Two-dimensional polyacrylamide gel electrophoresis and reverse transcriptase-PCR studies indicated that salicylate induced expression of the sdg cluster.

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“…Many C12O have been cloned and characterized from a variety of different bacteria, such as Acinetobacter , Arthrobacter , Corynebacterium , Pseudomonas , Rhodocococcus , Stenotrophomonas , and Streptomyces . Additionally, global microbial genome and environmental metagenome sequencing efforts are also contributing ever‐increasing genetic information to develop C12O.…”

Section: Introductionmentioning

confidence: 99%

Genetic diversity of catechol 1,2-dioxygenase in the fecal microbial metagenome

Xiong

Deng

et al. 2017

J. Basic Microbiol

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Catechol 1,2-dioxygenase is the key enzyme that catalyzes the cleavage of the aromatic ring of catechol. We explored the genetic diversity of catechol 1,2-dioxygenase in the fecal microbial metagenome by PCR with degenerate primers. A total of 35 gene fragments of C12O were retrieved from microbial DNA in the feces of pygmy loris. Based on phylogenetic analysis, most sequences were closely related to C12O sequences from Acinetobacter. A full-length C12O gene was directly cloned, heterologously expressed in Escherichia coli, and biochemically characterized. Purified catPL12 had optimum pH and temperature pH 8.0 and 25 °C and retained 31 and 50% of its maximum activity when assayed at 0 and 35 °C, respectively. The enzyme was stable at 25 and 37 °C, retaining 100% activity after pre-incubation for 1 h. The kinetic parameters of catPL12 were determined. The enzyme had apparent K of 67 µM, V of 7.3 U/mg, and k of 4.2 s for catechol, and the cleavage activities for 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol were much less than for catechol, and no activity with hydroquinone or protocatechuate was detected. This study is the first to report the molecular and biochemical characterizations of a cold-adapted catechol 1,2-dioxygenase from a fecal microbial metagenome.

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Cloning and expression of thermophilic catechol 1,2-dioxygenase gene (catA) from Streptomyces setonii

Cited by 7 publications

References 14 publications

An inducibleStreptomycesgene cluster involved in aromatic compound metabolism

An inducibleStreptomycesgene cluster involved in aromatic compound metabolism

Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46

Genetic diversity of catechol 1,2-dioxygenase in the fecal microbial metagenome

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