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Literature summary extracted from

  • Jenkins, R.J.; Dotson, G.D.
    A continuous fluorescent enzyme assay for early steps of lipid A biosynthesis (2012), Anal. Biochem., 425, 21-27.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.3.1.129 drug development Lpxa is a viable target for antimicrobial drug development Escherichia coli
2.3.1.191 drug development LpxD is a viable target for antimicrobial development Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.129 gene lpxA, cloning from strain strain MG1655, subcloning in strain XL-1 Blue, expression in Escherichia coli strain BL21-AI/pET24a Escherichia coli
2.3.1.191 gene lpxD, expression as His6-tagged protein in Escherichia coli strain BL21(DE3) Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.129 additional information
-
 additional information Michaelis-Menten steady-state kinetics of LpxA, overview Escherichia coli
2.3.1.191 0.004 0.012 UDP-3-O-[(3R)-3-hydroxymyristoyl]-alpha-D-glucosamine pH 8.0, 25°C, recombinant His6-tagged LpxD Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.129 an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine Escherichia coli
-
 an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
 ?
2.3.1.191 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-3-O-[(3R)-3-hydroxymyristoyl]-alpha-D-glucosamine Escherichia coli
-
 UDP-2,3-bis[O-(3R)-3-hydroxymyristoyl]-alpha-D-glucosamine + a holo-[acyl-carrier protein]
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.129 Escherichia coli
-
 strain MG1655, gene lpxA
-
2.3.1.191 Escherichia coli
-
 gene lpxD
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.129 recombinant LpxA from Echerichia coli strain BL21-AI/pET24a by Reactive Green 19 affinity chromatography, dialysis, and ion exchange chromatography Escherichia coli
2.3.1.191 recombinant His6-tagged LpxD from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.129 an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
-
 Escherichia coli an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
 ?
2.3.1.129 an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine assay is optimal at 10 nM LpxA Escherichia coli an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
 ?
2.3.1.191 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-3-O-[(3R)-3-hydroxymyristoyl]-alpha-D-glucosamine
-
 Escherichia coli UDP-2,3-bis[O-(3R)-3-hydroxymyristoyl]-alpha-D-glucosamine + a holo-[acyl-carrier protein]
-
 ir
2.3.1.191 additional information fluorescent enzyme assay, method optimization, overview Escherichia coli ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.129 LpxA
-
 Escherichia coli
2.3.1.129 type II ACP-dependent UDP-N-acetylglucosamine acyltransferase
-
 Escherichia coli
2.3.1.129 type II acyl carrier protein-dependent UDP-N-acetylglucosamine acyltransferase
-
 Escherichia coli
2.3.1.129 UDP-N-acetylglucosamine acyltransferase
-
 Escherichia coli
2.3.1.191 LpxD
-
 Escherichia coli
2.3.1.191 UDP-3-O-(R-3-hydroxyacyl)-glucosamine acyltransferase
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.129 25
-
 assay at Escherichia coli
2.3.1.191 25
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.191 3
-
 UDP-3-O-[(3R)-3-hydroxymyristoyl]-alpha-D-glucosamine pH 8.0, 25°C, recombinant His6-tagged LpxD Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.129 8
-
 assay at Escherichia coli
2.3.1.191 8
-
 assay at Escherichia coli

General Information

EC Number General Information Comment Organism
2.3.1.129 evolution the enzymes in early lipid A biosynthesis are highly conserved in gram-negative pathogens. LpxA and LpxD are structurally homologous possessing the unique left-handed beta-helix fold, which stems from an extensive hexapeptide repeat motif in their respective primary amino acid sequences Escherichia coli
2.3.1.129 metabolism UDP-N-acetylglucosamine acyltransferase, LpxA, catalyzes the first step of lipid A biosynthesis Escherichia coli
2.3.1.129 physiological function UDP-GlcNAc is acylated at the 3-hydroxyl through a thermodynamically unfavorable reaction catalyzed by the type II acyl carrier protein-dependent UDP-N-acetylglucosamine acyltransferase, LpxA, first step in biosynthesis of lipid A and essential for survival among Gram-negative bacteria that synthesize lipopolysaccharide. Lipopolysaccharide contains the glycolipid lipid A, which anchors it to the membrane through fatty acyl chains, a core polysaccharide region, and an O-antigen repeat. Lipid A is an essential moiety necessary for survival of the bacterium and further plays a crucial role in natural antibacterial resistance and bacterial sepsis Escherichia coli
2.3.1.191 metabolism LpxD catalyzes the third step of lipid A biosynthesis, an acyl-acyl carrier protein (ACP)-dependent transfer of a fatty acyl moiety to a UDP-glucosamine core ring, overview Escherichia coli
2.3.1.191 physiological function LpxD is essential for survival in Gram-negative bacteria Escherichia coli