β-Ketoacyl Acyl Carrier Protein Synthase III (FabH) Is Essential for Fatty Acid Biosynthesis in
            <i>Streptomyces coelicolor</i>
            A3(2)

Revill, W. Peter; Bibb, Maureen J.; Scheu, Ann-Karolin; Kieser, Helen J.; Hopwood, David A.

doi:10.1128/jb.183.11.3526-3530.2001

Cited by 68 publications

(48 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The fabH gene could be deleted only if a second copy of it was introduced into the cell. In this work and comparable experiments with E. coli, the additional copy of fabH was delivered as a chromosomal copy distant from its loci (11,16).…”

Section: Resultsmentioning

confidence: 99%

“…Recently, clear evidence has been obtained that FabH is an essential enzyme for fatty acid biosynthesis in E. coli, Streptomyces coelicolor, and Lactococcus lactis (11,16). In the cases of E. coli and S. coelicolor it was shown that the chromosomal fabH gene could be removed only if an additional copy of fabH was present; for S. coelicolor it was a second copy of the natural fabH gene (16), and in the case of E. coli it was the Salmonella enterica fabH (11). S. enterica is a close relative of E. coli, and it is known that the genes are fully interchangeable between the two species (11).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Alteration of the Fatty Acid Profile of Streptomyces coelicolor by Replacement of the Initiation Enzyme 3-Ketoacyl Acyl Carrier Protein Synthase III (FabH)

Florova

Reynolds

2005

J Bacteriol

View full text Add to dashboard Cite

The first elongation step of fatty acid biosynthesis by a type II dissociated fatty acid synthases is catalyzed by 3-ketoacyl-acyl carrier protein (ACP) synthase III (KASIII, FabH). This enzyme, encoded by the fabH gene, catalyzes a decarboxylative condensation between an acyl coenzyme A (CoA) primer and malonyl-ACP. In organisms such as Escherichia coli, which generate only straight-chain fatty acids (SCFAs), FabH has a substrate preference for acetyl-CoA. In streptomycetes and other organisms which produce a mixture of both SCFAs and branched-chain fatty acids (BCFAs), FabH has been shown to utilize straight-and branched-chain acyl-CoA substrates. We report herein the generation of a Streptomyces coelicolor mutant (YL/ecFabH) in which the chromosomal copy of the fabH gene has been replaced and the essential process of fatty acid biosynthesis is initiated by plasmid-based expression of the E. coli FabH (bearing only 35% amino acid identity to the Streptomyces enzyme). The YL/ecFabH mutant produces predominantly SCFAs (86%). In contrast, BCFAs predominate (ϳ70%) in both the S. coelicolor parental strain and S. coelicolor YL/sgFabH (a ⌬fabH mutant carrying a plasmid expressing the Streptomyces glaucescens FabH). These results provide the first unequivocal evidence that the substrate specificity of FabH observed in vitro is a determinant of the fatty acid made in an organism. The YL/ecFabH strain grows significantly slower on both solid and liquid media. The levels of FabH activity in cell extracts of YL/ecFabH were also significantly lower than those in cell extracts of YL/sgFabH, suggesting that a decreased rate of fatty acid synthesis may account for the observed decreased growth rate. The production of low levels of BCFAs in YL/ecFabH suggests either that the E. coli FabH is more tolerant of different acyl-CoAs substrates than previously thought or that there is an additional pathway for initiation of BCFA biosynthesis in Streptomyces coelicolor.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Alteration of the Fatty Acid Profile of Streptomyces coelicolor by Replacement of the Initiation Enzyme 3-Ketoacyl Acyl Carrier Protein Synthase III (FabH)

Florova

Reynolds

2005

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…In E. coli, FabH is subject to stringent feedback regulation by long-chain acyl-ACP so as to coordinate the rate of initiation with that of elongation (36). FabH is highly conserved, possessing an invariant triad of active site residues (28), as well as being essential across a diverse spectrum of both Gram-positive and Gram-negative bacteria that includes E. coli (47), Streptomyces coelicolor (67), and Lactococcus lactis (47). The uniqueness of the KASIII domain, conservation among FabH orthologs, and regulatory role in controlling FAS flux all suggest potential for broad-spectrum and bacterial selective inhibition.…”

Section: Trans-2-enoyl-acp (42 54) and Then Reduced To A Saturated mentioning

confidence: 99%

Pseudomonas aeruginosa Directly Shunts β-Oxidation Degradation Intermediates into De Novo Fatty Acid Biosynthesis

Yuan¹,

Leeds²,

Meredith³

2012

J Bacteriol

View full text Add to dashboard Cite

We identified the fatty acid synthesis (FAS) initiation enzyme in Pseudomonas aeruginosa as FabY, a β-ketoacyl synthase KASI/II domain-containing enzyme that condenses acetyl coenzyme A (acetyl-CoA) with malonyl-acyl carrier protein (ACP) to make the FAS primer β-acetoacetyl-ACP in the accompanying article (Y. Yuan, M. Sachdeva, J. A. Leeds, and T. C. Meredith, J. Bacteriol. 194:5171-5184, 2012). Herein, we show that growth defects stemming from deletion of fabY can be suppressed by supplementation of the growth media with exogenous decanoate fatty acid, suggesting a compensatory mechanism. Fatty acids eight carbons or longer rescue growth by generating acyl coenzyme A (acyl-CoA) thioester β-oxidation degradation intermediates that are shunted into FAS downstream of FabY. Using a set of perdeuterated fatty acid feeding experiments, we show that the open reading frame PA3286 in P. aeruginosa PAO1 intercepts C 8 -CoA by condensation with malonyl-ACP to make the FAS intermediate β-keto decanoyl-ACP. This key intermediate can then be extended to supply all of the cellular fatty acid needs, including both unsaturated and saturated fatty acids, along with the 3-hydroxyl fatty acid acyl groups of lipopolysaccharide. Heterologous PA3286 expression in Escherichia coli likewise established the fatty acid shunt, and characterization of recombinant β-keto acyl synthase enzyme activity confirmed in vitro substrate specificity for medium-chain-length acyl CoA thioester acceptors. The potential for the PA3286 shunt in P. aeruginosa to curtail the efficacy of inhibitors targeting FabY, an enzyme required for FAS initiation in the absence of exogenous fatty acids, is discussed.

show abstract

“…One of the essential enzymes, FabI, in the fatty acid biosynthetic pathway is a proven drug target with two marketed antibacterial agents, triclosan (antiseptic) and isoniazid (an anti-Mycobacterium tuberculosis agent) (12,13). The initiation condensing enzyme, FabH, and elongation condensing enzymes, FabF/B, are essential components of this biosynthetic pathway (14)(15)(16)(17) and consequently are highly conserved among key pathogens. Although no drugs targeting condensing enzymes are used in the clinic, two inhibitors, cerulenin (18) and thiolactomycin (19), with poor antibacterial activities were discovered more than two decades ago.…”

mentioning

confidence: 99%

Discovery of platencin, a dual FabF and FabH inhibitor with in vivo antibiotic properties

Wang

Kodali²,

Lee³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

371

366

View full text Add to dashboard Cite

Emergence of bacterial resistance is a major issue for all classes of antibiotics; therefore, the identification of new classes is critically needed. Recently we reported the discovery of platensimycin by screening natural product extracts using a target-based whole-cell strategy with antisense silencing technology in concert with cell free biochemical validations. Continued screening efforts led to the discovery of platencin, a novel natural product that is chemically and biologically related but different from platensimycin. Platencin exhibits a broad-spectrum Gram-positive antibacterial activity through inhibition of fatty acid biosynthesis. It does not exhibit cross-resistance to key antibiotic resistant strains tested, including methicillin-resistant Staphylococcus aureus, vancomycin-intermediate S. aureus, and vancomycin-resistant Enterococci. Platencin shows potent in vivo efficacy without any observed toxicity. It targets two essential proteins, ␤-ketoacyl-[acyl carrier protein (ACP)] synthase II (FabF) and III (FabH) with IC 50 values of 1.95 and 3.91 g/ml, respectively, whereas platensimycin targets only FabF (IC 50 ‫؍‬ 0.13 g/ml) in S. aureus, emphasizing the fact that more antibiotics with novel structures and new modes of action can be discovered by using this antisense differential sensitivity wholecell screening paradigm.platensimycin ͉ natural product ͉ thiolactomycin ͉ antisense ͉ fatty acid synthesis

show abstract

β-Ketoacyl Acyl Carrier Protein Synthase III (FabH) Is Essential for Fatty Acid Biosynthesis in Streptomyces coelicolor A3(2)

Cited by 68 publications

References 19 publications

Alteration of the Fatty Acid Profile of Streptomyces coelicolor by Replacement of the Initiation Enzyme 3-Ketoacyl Acyl Carrier Protein Synthase III (FabH)

Alteration of the Fatty Acid Profile of Streptomyces coelicolor by Replacement of the Initiation Enzyme 3-Ketoacyl Acyl Carrier Protein Synthase III (FabH)

Pseudomonas aeruginosa Directly Shunts β-Oxidation Degradation Intermediates into De Novo Fatty Acid Biosynthesis

Discovery of platencin, a dual FabF and FabH inhibitor with in vivo antibiotic properties

Contact Info

Product

Resources

About