Isolation and characterization of the beta-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12.

Tsay, Jiu-Tsair; Oh, W; Larson, Timothy J.; Jackowski, Suzanne; Rock, Charles O.

doi:10.1016/s0021-9258(19)50498-7

“…Bacterial fatty acid synthesis and the elongation of fatty acids can be initiated by FabH, one of the beta-ketoacyl-acyl-carrier protein synthases ( Cronan, Jr. et al, 2008 , Heath and Rock, 1996 , Tsay et al, 1992 ). In the development of antibacterial and antimicrobial drugs, FabI (enoyl-ACP reductase) and FabH have been targeted especially; isoniazid for tuberculosis operates in this way, as do cerulenin and thiolactomycin ( Banerjee et al, 1994 , Mdluli et al, 1998 , Moche et al, 1999 , Rozwarski et al, 1998 ).…”

Section: Discussionmentioning

confidence: 99%

Biochemical and in silico study of leaf and bark extracts from Aphanamixis polystachya against common pathogenic bacteria

Paul

¹

,

Mahmud

²

,

Hasan

³

et al. 2021

Saudi Journal of Biological Sciences

9

1

0

View full text Add to dashboard Cite

Aphanamixis polystachya may be a natural, renewable resource against antibiotic-resistant bacterial infections. The antibacterial activity of A. polystachya leaf and bark extracts was investigated against three antibiotic-resistant bacterial species and one fungus. Methanolic leaf extract showed only limited antibacterial activity but both methanolic and aqueous bark extract showed high antimicrobial activity. In an antioxidant activity test, leaf and bark extracts exhibited 50% free radical scavenging at a concentration of 107.14 ± 3.14 μg/mL and 97.13 ± 3.05 μg/mL, respectively, indicating that bark extracts offer more antioxidative activity than leaf extracts. Bark extracts also showed lower toxicity than leaf extracts. This suggests that bark extracts may offer greater development potential than leaf extracts. The molecular dynamics were also investigated through the simulated exploration of multiple potential interactions to understand the interaction dynamics (root-mean-square deviation, solvent-accessible surface area, radius of gyration, and the hydrogen bonding of chosen compounds to protein targets) and possible mechanisms of inhibition. This molecular modeling of compounds derived from A. polystachya revealed that inhibition may occur by binding to the active sites of the target proteins of the tested bacterial strains. A. polystachya bark extract may be used as a natural source of drugs to control antibiotic-resistant bacteria.

show abstract

“…Cys112 provides the essential thiol group in the enzymatic reaction of condensing enzymes [12,21,25]. In FabH, Cys112 is positioned at the bottom of a tunnel measuring 20 Å deep and 5 Å in diameter.…”

Section: The Active Sitementioning

confidence: 99%

“…The wild-type and mutant FabH proteins were assayed for condensation (β-ketobutyryl-ACP synthase) activities according to [11,12]. Briefly, 40 µl reactions were set up in 0.1 M sodium phosphate, pH 7.0, containing 25 µM ACP, 1 mM 2-mercaptoethanol, 65 µM malonyl-CoA, 45 µM acetyl-CoA (specific activity, 59 mCi/mmol; American Radiolabeled Chemicals, Inc.) and 0.2 µg FabD.…”

Section: Assays Of Fabh Proteinsmentioning

confidence: 99%

“…FabH or β-ketoacyl-ACP synthase III initiates the process, and FabB and FabF carry out the elongation reactions in subsequent cycles of fatty acid elongation. All three enzymes are thought to operate by a similar catalytic mechanism but FabH is distinguished from the others by its selectivity for acetyl-CoA over acyl-ACP [11,12]. FabH operates via a ping-pong mechanism utilizing an acetyl-enzyme intermediate (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The 1.8 Å crystal structure and active-site architecture of β-ketoacyl-acyl carrier protein synthase III (FabH) from Escherichia coli

Davies

¹

,

Heath

²

,

White

³

et al. 2000

View full text Add to dashboard Cite

show abstract

“…

KASIII catalyzes the condensation of acetyl-COA with malonyl-acyl carrier protein in dissociated (type 11) fatty acid synthase systems, such as those found in plant and bacterial cells. Initially identified as a cerulenin-resistant condensing enzyme (Jackowski and Rock, 1987;Jaworski et al 1989; Clough et al 1992), genes encoding this protein have been identified in Escherichiu coli (Tsay et al, 1992), a red alga (Reith 1993), and spinach (Tai and. Evidence that acetyl-COA is the major primer of fatty acid biosynthesis in spinach has recently been presented (Jaworski et al 1993), and a role for this enzyme in regulation of fatty acid biosynthesis in bacteria has been suggested (Jackowski and Rock, 1987).

…”

mentioning

confidence: 99%

cDNA Clones Encoding [beta]-Ketoacyl-Acyl Carrier Protein Synthase III from Cuphea wrightii

Slabaugh¹,

Tai²,

Jaworski³

et al. 1995

View full text Add to dashboard Cite

KASIII catalyzes the condensation of acetyl-COA with malonyl-acyl carrier protein in dissociated (type 11) fatty acid synthase systems, such as those found in plant and bacterial cells. Initially identified as a cerulenin-resistant condensing enzyme (Jackowski and Rock, 1987;Jaworski et al. 1989; Clough et al. 1992), genes encoding this protein have been identified in Escherichiu coli (Tsay et al., 1992), a red alga (Reith 1993), and spinach (Tai and. Evidence that acetyl-COA is the major primer of fatty acid biosynthesis in spinach has recently been presented (Jaworski et al. 1993), and a role for this enzyme in regulation of fatty acid biosynthesis in bacteria has been suggested (Jackowski and Rock, 1987). We are interested in whether the KASIII reaction represents a regulatory point in the rapid lipid biosynthesis observed in developing embryos of the oilseed plant Cupheu (Slabas et al., 1982). Here we report the sequence of two cDNA clones encoding KASIII in Cupkeu wrightii (Table I).A cDNA library in the A vector Uni-ZAPII was constructed from poly(A)+ RNA isolated from developing embryos of C. wrightii. A single clone was identified by relaxed-stringency screening of 120,000 plaques with a 600-bp segment of the spinach KASIII gene. The insert from this clone, cwKASIIIa, was used to rescreen the library, yielding a second clone, cwKASIIIb. DNA sequences were determined by the dideoxy method using an autosequencer (Applied Biosystems model 373A, version 12.0). Both clones contained a complete coding sequence.Preproteins of 400 and 402 amino acids are predicted from translation of the open reading frames of cDNA clones cwKASIIIa (1696 nucleotides) and cwKASIIIb (1827 nucleotides), respectively. The deduced amino acid sequences are 86% identical to each other, 71% identical to spinach KASIII (accession No. Z22771), and show 45, 37, and 36% homology to Porphyru umbiliculis chloroplast fabH This work was funded by grants from the U.S. Department of Agriculture (No. 58-5114-9-1002 and No. 91-37300-6569) and by the Procter and Gamble Company. Experiment station number 10,596 (Technical Paper Number).

show abstract

Isolation and characterization of the beta-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12.

Cited by 262 publications

References 52 publications

Biochemical and in silico study of leaf and bark extracts from Aphanamixis polystachya against common pathogenic bacteria

Biochemical and in silico study of leaf and bark extracts from Aphanamixis polystachya against common pathogenic bacteria

The 1.8 Å crystal structure and active-site architecture of β-ketoacyl-acyl carrier protein synthase III (FabH) from Escherichia coli

cDNA Clones Encoding [beta]-Ketoacyl-Acyl Carrier Protein Synthase III from Cuphea wrightii

Contact Info

Product

Resources

About