Degradation of Aromatics and Chloroaromatics by<i>Pseudomonas</i>sp. Strain B13: Cloning, Characterization, and Analysis of Sequences Encoding 3-Oxoadipate:Succinyl-Coenzyme A (CoA) Transferase and 3-Oxoadipyl-CoA Thiolase

Göbel, Markus; Kassel-Cati, Kerstin; Schmidt, Eberhard; Reineke, Walter

doi:10.1128/jb.184.1.216-223.2002

Cited by 28 publications

(16 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Accordingly, ␤-ketoadipate succinyl-CoA transferase activity may be encoded by either pca and/or cat genes (pcaIJ and catIJ, respectively), and these may share sequence similarity (15,25). Amino acid sequence identity between the S. meliloti PcaIJ and that of A. baylyi (25) (PcaI, 21%; PcaJ, 20%), P. putida (33) (PcaI, 21%; PcaJ, 23%), Bradyrhizobium japonicum (23) (PcaI, 17%; PcaJ, 20%), and even A. tumefaciens (53) (PcaI, 19%; PcaJ, 21%) is limited.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the β-Ketoadipate Pathway inSinorhizobium meliloti

MacLean

MacPherson

Aneja

et al. 2006

Appl Environ Microbiol

View full text Add to dashboard Cite

Aromatic compounds represent an important source of energy for soil-dwelling organisms. The ␤-ketoadipate pathway is a key metabolic pathway involved in the catabolism of the aromatic compounds protocatechuate and catechol, and here we show through enzymatic analysis and mutant analysis that genes required for growth and catabolism of protocatechuate in the soil-dwelling bacterium Sinorhizobium meliloti are organized on the pSymB megaplasmid in two transcriptional units designated pcaDCHGB and pcaIJF. The pcaD promoter was mapped by primer extension, and expression from this promoter is demonstrated to be regulated by the LysR-type protein PcaQ. ␤-Ketoadipate succinyl-coenzyme A (CoA) transferase activity in S. meliloti was shown to be encoded by SMb20587 and SMb20588, and these genes have been renamed pcaI and pcaJ, respectively. These genes are organized in an operon with a putative ␤-ketoadipyl-CoA thiolase gene (pcaF), and expression of the pcaIJF operon is shown to be regulated by an IclR-type transcriptional regulator, SMb20586, which we have named pcaR. We show that pcaR transcription is negatively autoregulated and that PcaR is a positive regulator of pcaIJF expression and is required for growth of S. meliloti on protocatechuate as the carbon source. The characterization of the protocatechuate catabolic pathway in S. meliloti offers an opportunity for comparison with related species, including Agrobacterium tumefaciens. Differences observed between S. meliloti and A. tumefaciens pcaIJ offer the first evidence of pca genes that may have been acquired after speciation in these closely related species.

show abstract

Section: Resultsmentioning

confidence: 99%

Characterization of the β-Ketoadipate Pathway inSinorhizobium meliloti

MacLean

MacPherson

Aneja

et al. 2006

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Several bacterial species metabolize carboxylates by ligating them to CoA (13,16,32,49). Ciprofibrate, which like GFZ is a carboxylate, is converted to ciprofibroyl-CoA in rat and marmoset livers (8,11).…”

Section: Vol 191 2009 Gfz Inhibits Bacterial Enoyl Reductases 5267mentioning

confidence: 99%

Gemfibrozil Inhibits Legionella pneumophila and Mycobacterium tuberculosis Enoyl Coenzyme A Reductases and Blocks Intracellular Growth of These Bacteria in Macrophages

Reich-Slotky¹,

Kabbash

Della‐Latta³

et al. 2009

J Bacteriol

View full text Add to dashboard Cite

We report here that gemfibrozil (GFZ) inhibits axenic and intracellular growth of Legionella pneumophila and of 27 strains of wild-type and multidrug-resistant Mycobacterium tuberculosis in bacteriological medium and in human and mouse macrophages, respectively. At a concentration of 0.4 mM, GFZ completely inhibited L. pneumophila fatty acid synthesis, while at 0.12 mM it promoted cytoplasmic accumulation of polyhydroxybutyrate. To assess the mechanism(s) of these effects, we cloned an L. pneumophila FabI enoyl reductase homolog that complemented for growth an Escherichia coli strain carrying a temperature-sensitive enoyl reductase and rendered the complemented E. coli strain sensitive to GFZ at the nonpermissive temperature. GFZ noncompetitively inhibited this L. pneumophila FabI homolog, as well as M. tuberculosis InhA and E. coli FabI.The advent of AIDS and the emergence of many multidrugresistant bacterial species have led to renewed efforts to find new antibiotics. The most commonly used antibiotics act by blocking bacterium-specific DNA, RNA, or protein synthesis. Mycobacterium tuberculosis is a major exception to this generalization. While streptomycin, an inhibitor of bacterial protein synthesis, was the first antibiotic to be used successfully to treat M. tuberculosis, isoniazid (INH), an inhibitor of mycobacterial lipid synthesis, is presently the drug most commonly used to treat infections with this organism (2, 43). The differential sensitivity to INH of M. tuberculosis versus mammalian cells reflects the fact that most bacterial fatty acid synthases (type II synthases) are comprised of discrete, separable enzymes encoded by separate genes, while mammalian fatty acid synthases (type I) are dimeric proteins in which a single polypeptide catalyzes the seven enzymatic activities of fatty acid synthesis (21, 52).In previous studies (45), we reported that gemfibrozil (GFZ), a commonly prescribed and well-tolerated hypolipidemic drug, inhibits the export of various organic anions, including penicillin and fluoroquinolones, from murine macrophages, thereby elevating the intracellular concentration of these antibiotics and enhancing their capacity to block intracellular growth of Listeria monocytogenes. In exploring this system, we discovered that while GFZ has no effect on axenic or intracellular growth of Listeria monocytogenes, it inhibits axenic growth of all Legionella pneumophila strains tested and of 5 wild-type and 22 multidrug-resistant strains of M. tuberculosis and inhibits intracellular growth of L. pneumophila Philadelphia-1 and M. tuberculosis H37RV in human and mouse macrophages, respectively.Both M. tuberculosis and L. pneumophila are facultative intracellular pathogens that enter host macrophages by phagocytosis (25, 26), grow in nonlysosomal membrane-bound cytoplasmic vacuoles (24), have special nutrient requirements (38, 54, 55), and produce a relatively unique spectrum of membrane lipids (7, 57). However, M. tuberculosis is a slow-growing and dangerous organism with which to work. In cont...

show abstract

“…YdiF is grouped with ϳ330 other proteins in the coenzyme A transferase superfamily IPR004165 (InterPro data base (30)) with rather diverse substrate specificities (31)(32)(33)(34)(35). Within the E. coli K12 genome, the individual N-terminal domain (residues 12-255) and C-terminal domain (residues 285-512) of YdiF are related in sequence to AtoD (24% identity) and AtoA (25% identity), representing the ␣-and ␤-subunits, respectively, of ACT (36).…”

Section: Ydif Is An Acyl-coa Transferasementioning

confidence: 99%

Crystallographic Trapping of the Glutamyl-CoA Thioester Intermediate of Family I CoA Transferases

Rangarajan

Ajamian

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Coenzyme A is a cofactor utilized by as many as 4% of all enzymes for a diverse variety of biological functions, including cell-cell-mediated recognition, nerve impulse conductance, transcription, and fatty acid biosynthesis and degradation (1, 2). Mainly, these reactions involve the binding and transfer of an acyl group from one substrate to another as part of an enzymatic reaction; it has been noted that coenzyme A is the most prominent acyl group carrier in all living systems (3). Enzymecatalyzed reactions employing CoA thioesters can be divided into two categories, (i) those where the thioester carbonyl C atom reacts as an electrophile and (ii) those where the thioester ␣-carbon is deprotonated and reacts as a nucleophile, in Claisen enzymes (1). CoA transferases, which catalyze the reversible transfer of CoA from a donor CoA thioester to a carboxylic acid acceptor generating the free donor and a new acyl-CoA (Scheme 1), belong to the first category of enzymes. Among the large number of CoA transferases, much attention has focused on mitochondrial succinyl-CoA:3-oxoacid CoA-transferase (SCOT), 4 as its autosomal recessive deficiency in humans results in improper ketone body utilization causing episodic severe ketosis, hypoglycemia, and ultimately coma (4, 5).Three classes of CoA transferases have been defined based mainly on mechanistic and sequence criteria (6). Family I enzymes employ as acceptors 3-oxoacids, short-chain fatty acids, or glutaconate. These enzymes operate with a ping-pong kinetic mechanism and form a covalent thioester intermediate (7). The most thoroughly studied member of the Family I CoA transferases is SCOT. Family II consists of the multifunctional enzymes citrate or citramalate lyase, and unlike Family I enzymes, they do not form a covalent thioester intermediate. Family III enzymes have been discovered more recently, and are distinct both mechanistically (6, 8) and structurally (9) A wealth of biochemical and mechanistic data are available for SCOT, largely based on the pioneering studies of Jencks and collaborators (7,(11)(12)(13). These studies established a landmark for the concept of substrate binding energy utilization by an enzyme to effect catalysis, showing that SCOT utilizes its covalent (␥-glutamyl-CoA thioester) and noncovalent interactions with the CoA moiety of the acyl-CoA substrate differentially to reduce the Gibbs activation energy required for catalysis (13). The utilization of this binding energy for catalysis differs for different chemical moieties within the CoA cofactor, as well for the different steps along the reaction coordinate. Although crystal structures are available for three Family I CoA transferases, including glutaconate CoA transferase (GCT) from Acidaminococcus fermentens (14), acetate-CoA transferase (ACT, ␣-subunit) from Escherichia coli (15), and SCOT from pig heart (16, 17), no structure has yet been determined with bound substrate or product. The absence of an enzyme-substrate co-crystal structure for any Family I CoA transferase has prevented...

show abstract

Degradation of Aromatics and Chloroaromatics byPseudomonassp. Strain B13: Cloning, Characterization, and Analysis of Sequences Encoding 3-Oxoadipate:Succinyl-Coenzyme A (CoA) Transferase and 3-Oxoadipyl-CoA Thiolase

Cited by 28 publications

References 55 publications

Characterization of the β-Ketoadipate Pathway inSinorhizobium meliloti

Characterization of the β-Ketoadipate Pathway inSinorhizobium meliloti

Gemfibrozil Inhibits Legionella pneumophila and Mycobacterium tuberculosis Enoyl Coenzyme A Reductases and Blocks Intracellular Growth of These Bacteria in Macrophages

Crystallographic Trapping of the Glutamyl-CoA Thioester Intermediate of Family I CoA Transferases

Contact Info

Product

Resources

About