Any feedback?
Literature summary extracted from
- Identification and partial characterization of a novel UDP-N-acetylenolpyruvoylglucosamine reductase/UDP-N-acetylmuramate L-alanine ligase fusion enzyme from Verrucomicrobium spinosum DSM 4136(T) (2016), Front. Microbiol., 7, 362 .
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 6.3.2.8 | drug development | the enzymes involved in synthesizing the bacterial cell wall are attractive targets for the design of antibacterial compounds, since this pathway is essential for bacteria and is absent in animals, particularly humans | Verrucomicrobium spinosum |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.3.1.98 | gene murB, Verrucomicrobium spinosum possesses a fusion open reading frame annotated by the locus tag (VspiD_010100018130). Recombinant expression in Escherichia coli strain Rosetta (DE3) pLysS. The ORF, which is predicted to encode the enzymes UDP-N-acetylenolpyruvoylglucosamine reductase (MurB) and UDP-N-acetylmuramate:L-alanine ligase (MurC) is cloned. In vivo analyses using functional complementation shows that the fusion gene is able to functionally complement Escherichia coli murB (mutant ST5 strain) and murC temperature sensitive mutants. The purified recombinant fusion enzyme (MurB/CVs) is endowed with UDP-N-acetylmuramate:L-alanine ligase activity. All attempts to demonstrate an in vitro UDP-N-acetylenolpyruvoylglucosamine reductase (MurB) activity are unsuccessful. Phylogenetic analysis reveals that this fusion enzyme can only be identified in specific lineages within the Verrucomicrobia phylum. The fusion gene is able to functionally complement two Escherichia coli strains that harbor mutations in the murB and murC genes | Verrucomicrobium spinosum |
| 6.3.2.8 | the ORF with the locus tag VspiD_010100018130, encodes the enzymes UDP-N-acetylenolpyruvoylglucosamine reductase (MurB) and UDP-N-acetylmuramate:L-alanine ligase (MurC), i.e. MurB/CVs, DNA and amino acid sequence determination and analysis, phylogenetic analysis, this fusion enzyme can only be identified in specific lineages within the Verrucomicrobia phylum. The fusion gene is able to functionally complement Escherichia coli murB and murC temperature-sensitive mutants, recombinant expression of His-tagged enzyme BurB/CVs in Escherichia coli strain Rosetta (DE3) pLysS | Verrucomicrobium spinosum |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 6.3.2.8 | 0.025 | - |
L-alanine | pH 9.0, 37°C, recombinnant enzyme MurB/CVs | Verrucomicrobium spinosum |  |
| 6.3.2.8 | 0.09 | - |
UDP-N-acetyl-alpha-D-muramate | pH 9.0, 37°C, recombinant enzyme MurB/CVs | Verrucomicrobium spinosum | |
| 6.3.2.8 | 0.47 | - |
ATP | pH 9.0, 37°C, recombinant enzyme MurB/CVs | Verrucomicrobium spinosum | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 1.3.1.98 | cytoplasm | - |
Verrucomicrobium spinosum | 5737 | - |
| 6.3.2.8 | cytoplasm | - |
Verrucomicrobium spinosum | 5737 | - |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 6.3.2.8 | Mg2+ | required, optimal at 10 mM for the UDP-N-acetylmuramate:L-alanine ligase activity of MurB/CVs | Verrucomicrobium spinosum | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.3.1.98 | UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine + NADPH + H+ | Verrucomicrobium spinosum | the enzyme MurB catalyzes the reduction of the enolpyruvyl moiety of UDP-GlcNAc-enolpyruvate to lactyl ether to produce UDP-N-acetylmuramic acid | UDP-N-acetyl-alpha-D-muramate + NADP+ | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine | Verrucomicrobium spinosum | - |
ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine | Verrucomicrobium spinosum DSM 4136 | - |
ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.3.1.98 | Verrucomicrobium spinosum | - |
- |
- |
| 1.3.1.98 | Verrucomicrobium spinosum DSM 4136 | - |
- |
- |
| 6.3.2.8 | Verrucomicrobium spinosum | - |
- |
- |
| 6.3.2.8 | Verrucomicrobium spinosum DSM 4136 | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 6.3.2.8 | recombinant His-tagged enzyme BurB/CVs from Escherichia coli strain Rosetta (DE3) pLysS by nickel affinity chromatography, diaylsis, and ultrafiltration | Verrucomicrobium spinosum |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.3.1.98 | UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine + NADPH + H+ | - |
Verrucomicrobium spinosum | UDP-N-acetyl-alpha-D-muramate + NADP+ | - |
? | |
| 1.3.1.98 | UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine + NADPH + H+ | the enzyme MurB catalyzes the reduction of the enolpyruvyl moiety of UDP-GlcNAc-enolpyruvate to lactyl ether to produce UDP-N-acetylmuramic acid | Verrucomicrobium spinosum | UDP-N-acetyl-alpha-D-muramate + NADP+ | - |
? | |
| 1.3.1.98 | UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine + NADPH + H+ | - |
Verrucomicrobium spinosum DSM 4136 | UDP-N-acetyl-alpha-D-muramate + NADP+ | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + glycine | reduced activity with glycine compared to L-alanine | Verrucomicrobium spinosum | ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-glycine | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + glycine | reduced activity with glycine compared to L-alanine | Verrucomicrobium spinosum DSM 4136 | ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-glycine | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine | - |
Verrucomicrobium spinosum | ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine | - |
Verrucomicrobium spinosum DSM 4136 | ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + L-serine | reduced activity with L-serine compared to L-alanine | Verrucomicrobium spinosum | ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-serine | - |
? | |
| 6.3.2.8 | ATP + UDP-N-acetyl-alpha-D-muramate + L-serine | reduced activity with L-serine compared to L-alanine | Verrucomicrobium spinosum DSM 4136 | ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-serine | - |
? | |
| 6.3.2.8 | additional information | in contrast to the UDP-N-acetylmuramate:L-alanine ligase (MurC) activity, the activity of the UDP-N-acetylenolpyruvoylglucosamine reductase (MurB, EC 1.3.1.98) cannot be detected in vitro | Verrucomicrobium spinosum | ? | - |
? | |
| 6.3.2.8 | additional information | in contrast to the UDP-N-acetylmuramate:L-alanine ligase (MurC) activity, the activity of the UDP-N-acetylenolpyruvoylglucosamine reductase (MurB, EC 1.3.1.98) cannot be detected in vitro | Verrucomicrobium spinosum DSM 4136 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.3.1.98 | ? | x * 87300, recombinant MurB/CVs fusion enzyme, SDS-PAGE | Verrucomicrobium spinosum |
| 1.3.1.98 | More | domain mapping of the MurB/CVs fusion enzyme of Verrucomicrobium spinosum and the individual MurB and MurC enzymes from Escherichia coli | Verrucomicrobium spinosum |
| 6.3.2.8 | More | enzyme domain organization, overview | Verrucomicrobium spinosum |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.3.1.98 | MurB | - |
Verrucomicrobium spinosum |
| 1.3.1.98 | MurBAb | - |
Verrucomicrobium spinosum |
| 1.3.1.98 | UDP-N-acetylenolpyruvoylglucosamine reductase | - |
Verrucomicrobium spinosum |
| 1.3.1.98 | VspiD_010100018130 | locus name | Verrucomicrobium spinosum |
| 6.3.2.8 | MurB/CVs | - |
Verrucomicrobium spinosum |
| 6.3.2.8 | UDP-N-acetylenolpyruvoylglucosamine reductase/UDP-N-acetylmuramate: L-alanine ligase | - |
Verrucomicrobium spinosum |
| 6.3.2.8 | UDP-N-acetylmuramate:L-alanine ligase | - |
Verrucomicrobium spinosum |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 6.3.2.8 | 44 | 46 | UDP-N-acetylmuramate:L-alanine ligase activity of MurB/CVs | Verrucomicrobium spinosum |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 6.3.2.8 | 9 | - |
UDP-N-acetylmuramate:L-alanine ligase activity of MurB/CVs | Verrucomicrobium spinosum |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.3.1.98 | NADPH | - |
Verrucomicrobium spinosum | |
| 6.3.2.8 | ATP | - |
Verrucomicrobium spinosum | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.3.1.98 | evolution | Verrucomicrobium spinosum possesses a fusion open reading frame annotated by the locus tag (VspiD_010100018130). The ORF, which is predicted to encode the enzymes UDP-N-acetylenolpyruvoylglucosamine reductase (MurB) and UDP-N-acetylmuramate:L-alanine ligase (MurC) is cloned. In vivo analyses using functional complementation shows that the fusion gene is able to complement Escherichia coli murB and murC temperature sensitive mutants. The purified recombinant fusion enzyme (MurB/CVs) is endowed with UDP-N-acetylmuramate:L-alanine ligase activity. All attempts to demonstrate an in vitro UDP-N-acetylenolpyruvoylglucosamine reductase (MurB) activity are unsuccessful. Phylogenetic analysis reveals that this fusion enzyme can only be identified in specific lineages within the Verrucomicrobia phylum | Verrucomicrobium spinosum |
| 1.3.1.98 | metabolism | MurB is involved in cytoplasmic steps of peptidoglycan biosynthesis the peptidoglycan biosynthesis pathway, overview | Verrucomicrobium spinosum |
| 6.3.2.8 | evolution | the ORF with the locus tag VspiD_010100018130, encodes the enzymes UDP-N-acetylenolpyruvoylglucosamine reductase (MurB, EC 1.3.1.98) and UDP-N-acetylmuramate:L-alanine ligase (MurC), i.e. MurB/CVs, DNA and amino acid sequence determination and analysis, this fusion enzyme can only be identified in specific lineages within the Verrucomicrobia phylum. Unusual MurB and MurC composition is prevalent in currently sequenced members of Verrucomicrobia | Verrucomicrobium spinosum |
| 6.3.2.8 | metabolism | the enzyme MurB/CVs is involved in the cytoplasmic steps of peptidoglycan biosynthesis catalyzing the second and third step of the pathway, overview | Verrucomicrobium spinosum |
| 6.3.2.8 | physiological function | the fusion gene encoding MurB/CVs is able to complement Escherichia coli murB and murC temperature-sensitive mutants | Verrucomicrobium spinosum |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
