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Literature summary extracted from

  • Al-Dabbagh, B.; Olatunji, S.; Crouvoisier, M.; El Ghachi, M.; Blanot, D.; Mengin-Lecreulx, D.; Bouhss, A.
    Catalytic mechanism of MraY and WecA, two paralogues of the polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase superfamily (2016), Biochimie, 127, 249-257.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.7.8.13 drug development the specificity of the enzyme to eubacteria makes it a promising potential target to be exploited for antibacterial discovery to combat emerging resistance Bacillus subtilis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.8.13 gene mraY, cloning in Escherichia coli strain DH5alpha Bacillus subtilis
2.7.8.33 gene wecA, cloning in Escherichia coli strain DH5alpha Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
2.7.8.13 D98A site-directed mutagenesis, the mutant shows altered pH dependence compared to the wild-type enzyme Bacillus subtilis
2.7.8.33 D72A site-directed mutagenesis, very low activity of the mutant protein at pH 8.0, which represents only about 1.2% of the wild-type activity. At pH 7.0, the mutant protein is totally inactive. Increasing the pH from 8 to 9 results in a 2.5fold increase of the D72A mutant activity, while the wild-type enzyme activity rather decreases, from 310 to 240 U/mg of protein Thermotoga maritima

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.8.13 dodecylamine a competitive inhibitor of MraY Bacillus subtilis
2.7.8.13 additional information no inhibition by 1-diphospho-N-acetylmuramoyl-pentapeptide Bacillus subtilis
2.7.8.33 dodecylamine
-
 Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.8.13 additional information
-
 additional information the enzyme exhibits Michaelis-Menten kinetics towards the heptaprenyl phosphate and dodecaprenyl phosphate lipid substrates. The Km values for dodecaprenyl phosphate and heptaprenyl phosphate are 0.21 mM and 0.15 mM, respectively. The Km values are similar to that of the physiological lipid substrate, undecaprenyl phosphate with 0.16 mM Bacillus subtilis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.8.13 cell wall
-
 Bacillus subtilis 5618
-
2.7.8.33 cell wall
-
 Thermotoga maritima 5618
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.8.13 Mg2+ essentially required Bacillus subtilis
2.7.8.33 Mg2+ essentially required Thermotoga maritima

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate Bacillus subtilis
-
 UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate Bacillus subtilis 168
-
 UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 r
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate Thermotoga maritima the enzyme is highly specific for UDP-GlcNAc, its physiological substrate UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
 r
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate Thermotoga maritima ATCC 43589 the enzyme is highly specific for UDP-GlcNAc, its physiological substrate UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.8.13 Bacillus subtilis Q03521 gene mraY
-
2.7.8.13 Bacillus subtilis 168 Q03521 gene mraY
-
2.7.8.33 Thermotoga maritima Q9X1N5 gene wecA
-
2.7.8.33 Thermotoga maritima ATCC 43589 Q9X1N5 gene wecA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.8.33 recombinant wild-type and D72A mutant enzymes Thermotoga maritima

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate = UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol a one-step, single displacement mechanism. The oxyanion of the polyprenyl-phosphate attacks the b-phosphate of the nucleotide substrate, leading to the formation of product lipid I and liberation of UMP. The involvement of an invariant aspartyl residue in the deprotonation of the lipid substrate is possible. Formation of a covalent reaction intermediate Bacillus subtilis
a
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate = UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol a one-step, single displacement mechanism. The oxyanion of the polyprenyl-phosphate attacks the beta-phosphate of the nucleotide substrate, leading to the formation of lipid product and the liberation of UMP. The involvement of an invariant aspartyl residue in the deprotonation of the lipid substrate is possible Thermotoga maritima
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.8.13 additional information the enzyme does not display any diphosphatase activity on the nucleotide substrate. No activity with UDP-GlcNAc and 1-diphospho-MurNAc-pentapeptide. Enzyme catalytic mechanism and substrate specificity, overview. The essential aspartate residue, that is invariant in the superfamily, is Asp98 in Bacillus subtilis MraY, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. UDPMurNAc, UDP-MurNAc-L-Ala, UDP-MurNAc-L-Ala-D-Glu and UDPMurNAc-L-Ala-gamma-D-Glu-meso-diaminopimelate precursors of the peptidoglycan biosynthesis pathway are all substrates of the enzyme, no activity with 1-diphospho-MurNAc-pentapeptide. No activity with isopentenyl phosphate as lipid substrate, the enzyme is highly active when tested with neryl phosphate, heptaprenyl phosphate, dodecaprenyl phosphate, and pentadecaprenyl phosphate Bacillus subtilis ?
-
 ?
2.7.8.13 additional information the enzyme does not display any diphosphatase activity on the nucleotide substrate. No activity with UDP-GlcNAc and 1-diphospho-MurNAc-pentapeptide. Enzyme catalytic mechanism and substrate specificity, overview. The essential aspartate residue, that is invariant in the superfamily, is Asp98 in Bacillus subtilis MraY, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. UDPMurNAc, UDP-MurNAc-L-Ala, UDP-MurNAc-L-Ala-D-Glu and UDPMurNAc-L-Ala-gamma-D-Glu-meso-diaminopimelate precursors of the peptidoglycan biosynthesis pathway are all substrates of the enzyme, no activity with 1-diphospho-MurNAc-pentapeptide. No activity with isopentenyl phosphate as lipid substrate, the enzyme is highly active when tested with neryl phosphate, heptaprenyl phosphate, dodecaprenyl phosphate, and pentadecaprenyl phosphate Bacillus subtilis 168 ?
-
 ?
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate
-
 Bacillus subtilis UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate the forward and reverse exchange reactions require the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. No activity is detected with isopentenyl phosphate, but the enzyme is highly active with neryl phosphate, heptaprenyl phosphate, dodecaprenyl phosphate, and pentadecaprenyl phosphate Bacillus subtilis UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate
-
 Bacillus subtilis 168 UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) + undecaprenyl phosphate the forward and reverse exchange reactions require the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. No activity is detected with isopentenyl phosphate, but the enzyme is highly active with neryl phosphate, heptaprenyl phosphate, dodecaprenyl phosphate, and pentadecaprenyl phosphate Bacillus subtilis 168 UMP + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-MurNAc + undecaprenyl phosphate 24% activity compared to UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) Bacillus subtilis UMP + MurNAc-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-MurNAc + undecaprenyl phosphate 24% activity compared to UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) Bacillus subtilis 168 UMP + MurNAc-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-MurNAc-(oyl-L-Ala) + undecaprenyl phosphate 9% activity compared to UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) Bacillus subtilis UMP + MurNAc(oyl-L-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-MurNAc-(oyl-L-Ala) + undecaprenyl phosphate 9% activity compared to UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) Bacillus subtilis 168 UMP + MurNAc(oyl-L-Ala)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-MurNAc-(oyl-L-Ala-gamma-D-Glu) + undecaprenyl phosphate 36% activity compared to UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) Bacillus subtilis UMP + MurNAc(oyl-L-Ala-gamma-D-Glu)-diphosphoundecaprenol
-
 r
2.7.8.13 UDP-MurNAc-(oyl-L-Ala-gamma-D-Glu-meso-diaminopimelate) + undecaprenyl phosphate 51% activity compared to UDP-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala) Bacillus subtilis UMP + MurNAc(oyl-L-Ala-gamma-D-Glu-meso-diaminopimelate)-diphosphoundecaprenol
-
 r
2.7.8.33 additional information the enzyme does not display any diphosphatase activity on the nucleotide substrate. Enzyme catalytic mechanism and substrate specificity, overview. The minimal length of the carbon chain of the lipid substrate for an efficient catalysis is 35. The essential aspartate residue, that is invariant in the superfamily, is Asp72 in Thermotoga maritima WecA, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. No activity by the enzyme with UDP-galactose, UDP-GalNAc, GDP-glucose, ADP-ribose, UDP-glucuronic acid, GDP-D-mannose, and UDP-hexanolamine Thermotoga maritima ?
-
 ?
2.7.8.33 additional information the enzyme does not display any diphosphatase activity on the nucleotide substrate. Enzyme catalytic mechanism and substrate specificity, overview. The minimal length of the carbon chain of the lipid substrate for an efficient catalysis is 35. The essential aspartate residue, that is invariant in the superfamily, is Asp72 in Thermotoga maritima WecA, the residue is involved in the deprotonation of the lipid substrate during the catalytic process. No activity by the enzyme with UDP-galactose, UDP-GalNAc, GDP-glucose, ADP-ribose, UDP-glucuronic acid, GDP-D-mannose, and UDP-hexanolamine Thermotoga maritima ATCC 43589 ?
-
 ?
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate the enzyme is highly specific for UDP-GlcNAc, its physiological substrate Thermotoga maritima UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
 r
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate the forward and reverse exchange reactions required the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. The nucleotide substrate UDPMurNAc-pentapeptide, as well as the nucleotide product UMP, can bind to MraY in the absence of lipid ligands. The enzyme is highly specific for UDP-GlcNAc, its physiological substrate Thermotoga maritima UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
 r
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate the enzyme is highly specific for UDP-GlcNAc, its physiological substrate Thermotoga maritima ATCC 43589 UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
 r
2.7.8.33 UDP-N-acetyl-alpha-D-glucosamine + ditrans,octacis-undecaprenyl phosphate the forward and reverse exchange reactions required the presence of the second substrate, undecaprenyl phosphate and UMP, respectively. The nucleotide substrate UDPMurNAc-pentapeptide, as well as the nucleotide product UMP, can bind to MraY in the absence of lipid ligands. The enzyme is highly specific for UDP-GlcNAc, its physiological substrate Thermotoga maritima ATCC 43589 UMP + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.7.8.13 MraY
-
 Bacillus subtilis
2.7.8.13 polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase
-
 Bacillus subtilis
2.7.8.33 WecA
-
 Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.8.13 37
-
 assay at Bacillus subtilis
2.7.8.33 65
-
 assay at Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.8.13 additional information
-
 additional information the enzyme exhibits Michaelis-Menten kinetics towards the heptaprenyl phosphate and dodecaprenyl phosphate lipid substrates. The catalytic constants kcat are 295/min and 54/min, respectively. The kcat for heptaprenyl phosphate is by 6fold lower than those determined for the longer-chain length lipids undecaprenyl phosphate and dodecaprenyl phosphate with 340/min and 295/min, respectively Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.8.13 7.6
-
 assay at Bacillus subtilis
2.7.8.33 8
-
 assay at Thermotoga maritima

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.7.8.13 1
-
 pH 7.6, 37°C Bacillus subtilis dodecylamine

General Information

EC Number General Information Comment Organism
2.7.8.13 evolution the enzyme is a member of the polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase, P2HPT, superfamily Bacillus subtilis
2.7.8.13 physiological function enzyme MraY transferase catalyzes the first membrane step of bacterial cell wall peptidoglycan biosynthesis, namely the transfer of the N-acetylmuramoyl-pentapeptide moiety of the cytoplasmic precursor UDP- N-acetylmuramoyl-pentapeptide to the membrane transporter undecaprenyl phosphate (C55-P), yielding undecaprenyl-N-acetylmuramoyl-pentapeptide, i.e. lipid I. The MraY transferase initiates the membrane steps of peptidoglycan biosynthesis and is an essential enzyme for bacterial viability Bacillus subtilis
2.7.8.33 evolution the enzyme is a member of the polyprenyl-phosphate N-acetylhexosamine 1-phosphate transferase, P2HPT, superfamily Thermotoga maritima
2.7.8.33 metabolism WecA catalyzes the first membrane step of the biosynthesis of many cell wall polymers such as the O-antigen, teichoic acids and arabinogalactan Thermotoga maritima
2.7.8.33 physiological function enzyme WecA, catalyzes the transfer of the phospho-GlcNAc moiety of UDP-N-acetylglucosamine onto the same lipid carrier, leading to the formation of N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol that is essential for the synthesis of various bacterial cell envelope components. Undecaprenyl diphosphoryl-N-acetylglucosamine (lipid intermediate I) is a ubiquitous compound in all kingdoms of life. the enzyme is involved in the initiation of enterobacterial common antigen biosynthesis Thermotoga maritima