scholarly journals PhcS Represses Gratuitous Expression of Phenol-Metabolizing Enzymes in Comamonas testosteroni R5

2001 ◽  
Vol 183 (14) ◽  
pp. 4227-4234 ◽  
Author(s):  
Maki Teramoto ◽  
Shigeaki Harayama ◽  
Kazuya Watanabe
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Chemostat Culture ◽  
Transcriptional Regulators ◽  
Open Reading Frame ◽  
Comamonas Testosteroni ◽  
Metabolizing Enzymes ◽  
Unknown Mechanism ◽  
Reading Frame ◽  
Multicomponent Phenol Hydroxylase ◽  
Gntr Family

ABSTRACT We identified an open reading frame, designatedphcS, downstream of the transcriptional activator gene (phcR) for the expression of multicomponent phenol hydroxylase (mPH) in Comamonas testosteroni R5. The deduced product of phcS was homologous to AphS ofC. testosteroni TA441, which belongs to the GntR family of transcriptional regulators. The transformation of Pseudomonas aeruginosa PAO1c (phenol negative, catechol positive) with pROR502 containingphcR and the mPH genes conferred the ability to grow on phenol, while transformation with pROR504 containingphcS, phcR, and mPH genes did not confer this ability. The disruption of phcS in strain R5 had no effect on its phenol-oxygenating activity in a chemostat culture with phenol. The phenol-oxygenating activity was not expressed in strain R5 grown in a chemostat with acetate. In contrast, the phenol-oxygenating activity in the strain with a knockoutphcS gene when grown in a chemostat with acetate as the limiting growth factor was 66% of that obtained in phenol-grown cells of the strain with a knockout in the phcSgene. The disruption of phcS and/orphcR and the complementation in trans of these defects confirm that PhcS is a trans-acting repressor and that the unfavorable expression of mPH in thephcS knockout cells grown on acetate requires PhcR. These results show that the PhcS protein repressed the gratuitous expression of phenol-metabolizing enzymes in the absence of the genuine substrate and that strain R5 acted by an unknown mechanism in which the PhcS-mediated repression was overcome in the presence of the pathway substrate.

scholarly journals An AraC/XylS Family Member at a High Level in a Hierarchy of Regulators for Phenol-Metabolizing Enzymes in Comamonas testosteroni R5

2002 ◽  
Vol 184 (14) ◽  
pp. 3941-3946 ◽  
Author(s):  
Maki Teramoto ◽  
Kouhei Ohnishi ◽  
Shigeaki Harayama ◽  
Kazuya Watanabe
Keyword(s):  
Family Member ◽  
Bacterial Strain ◽  
Transcriptional Regulator ◽  
Transcriptional Regulators ◽  
Comamonas Testosteroni ◽  
Metabolizing Enzymes ◽  
Operon Expression ◽  
High Level ◽  
Expression Strain ◽  
Multicomponent Phenol Hydroxylase

ABSTRACT Comamonas testosteroni strain R5 expresses a higher level of phenol-oxygenating activity than any other bacterial strain so far characterized. The expression of the operon encoding multicomponent phenol hydroxylase (mPH), which is responsible for the phenol-oxygenating activity, is controlled by two transcriptional regulators, PhcS and PhcR, in strain R5. In this study, we identified a third transcriptional regulator for the mPH operon (PhcT) that belongs to the AraC/XylS family. While the disruption of phcT in strain R5 significantly reduced the expression of the mPH operon, it did not eliminate the expression. However, the disruption of phcT in strain R5 increased the expression of phcR. The phenol-oxygenating activity was abolished by the disruption of phcR, indicating that PhcT alone was not sufficient to activate the expression of the mPH operon. The disruption of phcS has been shown in our previous study to confer the ability of strain R5 to express the mPH operon in the absence of the genuine substrate for mPH. PhcT was not involved in the gratuitous expression. Strain R5 thus possesses a more elaborate mechanism for regulating the mPH operon expression than has been found in other bacteria.

scholarly journals A Novel Lipolytic Enzyme Located in the Outer Membrane of Pseudomonas aeruginosa

1999 ◽  
Vol 181 (22) ◽  
pp. 6977-6986 ◽  
Author(s):  
Susanne Wilhelm ◽  
Jan Tommassen ◽  
Karl-Erich Jaeger
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Outer Membrane ◽  
Open Reading Frame ◽  
Lipolytic Enzyme ◽  
Cell Fractionation ◽  
Sequence Alignments ◽  
Reading Frame ◽  
Amino Terminal

ABSTRACT A lipase-negative deletion mutant of Pseudomonas aeruginosa PAO1 still showed extracellular lipolytic activity toward short-chain p-nitrophenylesters. By screening a genomic DNA library of P. aeruginosa PAO1, an esterase gene, estA, was identified, cloned, and sequenced, revealing an open reading frame of 1,941 bp. The product ofestA is a 69.5-kDa protein, which is probably processed by removal of an N-terminal signal peptide to yield a 67-kDa mature protein. A molecular mass of 66 kDa was determined for35S-labeled EstA by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The amino acid sequence of EstA indicated that the esterase is a member of a novel GDSL family of lipolytic enzymes. The estA gene showed high similarity to an open reading frame of unknown function located in thetrpE-trpG region of P. putida and to a gene encoding an outer membrane esterase of Salmonella typhimurium. Amino acid sequence alignments led us to predict that this esterase is an autotransporter protein which possesses a carboxy-terminal β-barrel domain, allowing the secretion of the amino-terminal passenger domain harboring the catalytic activity. Expression of estA in P. aeruginosa andEscherichia coli and subsequent cell fractionation revealed that the enzyme was associated with the cellular membranes. Trypsin treatment of whole cells released a significant amount of esterase, indicating that the enzyme was located in the outer membrane with the catalytic domain exposed to the surface. To our knowledge, this esterase is unique in that it exemplifies in P. aeruginosa(i) the first enzyme identified in the outer membrane and (ii) the first example of a type IV secretion mechanism.

scholarly journals Glycine Betaine Transmethylase Mutant of Pseudomonas aeruginosa

2002 ◽  
Vol 184 (15) ◽  
pp. 4301-4303 ◽  
Author(s):  
Ana L. Serra ◽  
Javier F. Mariscotti ◽  
José L. Barra ◽  
Gloria I. Lucchesi ◽  
Carlos E. Domenech ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Sequence Analysis ◽  
Glycine Betaine ◽  
Nitrogen Sources ◽  
Open Reading Frame ◽  
Carbon And Nitrogen Sources ◽  
Reading Frame ◽  
Carbon And Nitrogen ◽  
Molecular Approaches ◽  
Knockout Gene

ABSTRACT The gene for glycine betaine transmethylase (gbt) was identified in Pseudomonas aeruginosa strain Fildes III by biochemical, physiological, and molecular approaches. Based on sequence analysis, the knockout gene corresponded to an open reading frame (ORF) named PA3082 in the genome of P. aeruginosa PAO1. The translated product of this ORF displayed similarity to transferases of different microorganisms. Mutation in gbt blocked the utilization of choline and glycine betaine as carbon and nitrogen sources.

scholarly journals Cloning, sequencing and analysis of the structural gene and regulatory region of the Pseudomonas aeruginosa chromosomal ampCβ-lactamase

1990 ◽  
Vol 272 (3) ◽  
pp. 627-631 ◽  
Author(s):  
J M Lodge ◽  
S D Minchin ◽  
L J V Piddock ◽  
S J W Busby
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Binding Site ◽  
Regulatory Protein ◽  
Regulatory Region ◽  
Open Reading Frame ◽  
Beta Lactamase ◽  
Chromosomal Gene ◽  
Reading Frame ◽  
Beta Lactamases ◽  
Lactamase Gene

The chromosomal gene from Pseudomonas aeruginosa encoding beta-lactamase has been cloned, and the sequence determined and compared with corresponding sequences of beta-lactamases from members of the enterobacteriaceae. Upstream of the beta-lactamase gene is an open reading frame which we postulate encodes a regulatory protein, AmpR. We identified a helix-turn-helix region in AmpR and a putative AmpR-binding site.

scholarly journals Identification of the Pseudomonas aeruginosa glmM Gene, Encoding Phosphoglucosamine Mutase

2000 ◽  
Vol 182 (16) ◽  
pp. 4453-4457 ◽  
Author(s):  
Isabel M. Tavares ◽  
Laure Jolly ◽  
Frédérique Pompeo ◽  
Jorge H. Leitão ◽  
Arsénio M. Fialho ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Open Reading Frame ◽  
Expression Vectors ◽  
Gene Encoding ◽  
Genome Database ◽  
Reading Frame ◽  
Peptidoglycan Biosynthesis ◽  
Essential Step ◽  
Residue Polypeptide

ABSTRACT A search for a potential algC homologue within thePseudomonas aeruginosa PAO1 genome database has revealed an open reading frame (ORF) of unknown function, ORF540 in contig 54 (July 1999 Pseudomonas genome release), that theoretically coded for a 445-amino-acid-residue polypeptide (I. M. Tavares, J. H. Leitão, A. M. Fialho, and I. Sá-Correia, Res. Microbiol. 150:105–116, 1999). The product of this gene is here identified as the phosphoglucosamine mutase (GlmM) which catalyzes the conversion of glucosamine-6-phosphate to glucosamine-1-phosphate, an essential step in the formation of the cell wall precursor UDP-N-acetylglucosamine. The P. aeruginosa gene has been cloned into expression vectors and shown to restore normal peptidoglycan biosynthesis and cell growth of a glmM Escherichia coli mutant strain. The GlmM enzyme from P. aeruginosa has been overproduced to high levels and purified to homogeneity in a six-histidine-tagged form. Beside its phosphoglucosamine mutase activity, the P. aeruginosaenzyme is shown to exhibit phosphomannomutase and phosphoglucomutase activities, which represent about 20 and 2% of its GlmM activity, respectively.

scholarly journals Molecular Analysis of Metallo-β-Lactamase Gene blaSPM-1-Surrounding Sequences from Disseminated Pseudomonas aeruginosa Isolates in Recife, Brazil

2004 ◽  
Vol 48 (4) ◽  
pp. 1406-1409 ◽  
Author(s):  
Laurent Poirel ◽  
Marcelo Magalhaes ◽  
Miguel Lopes ◽  
Patrice Nordmann
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Molecular Analysis ◽  
Open Reading Frame ◽  
Common Region ◽  
Reading Frame ◽  
Carbapenem Resistant ◽  
Lactamase Gene

ABSTRACT The spread of clonally related carbapenem-resistant Pseudomonas aeruginosa producing the metallo-β-lactamase SPM-1 was found in Recife, Brazil. Upstream of bla SPM-1, a novel common region (CR4) was identified, comprising an open reading frame, orf495, whose product shares significant identity with putative recombinases, such as Orf513. CR4 may be responsible for mobilization and expression of bla SPM-1.

Sign in / Sign up

Export Citation Format

Share Document