Literature summary for 6.3.2.8 extracted from

Naqvi, K.F.; Patin, D.; Wheatley, M.S.; Savka, M.A.; Dobson, R.C.; Gan, H.M.; Barreteau, H.; Blanot, D.; Mengin-Lecreulx, D.; Hudson, A.O.
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

Application	Comment	Organism
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)

Cloned (Comment)	Organism
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

Cloned (Comment)

Organism

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]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.025	-	L-alanine	pH 9.0, 37°C, recombinnant enzyme MurB/CVs	Verrucomicrobium spinosum
0.09	-	UDP-N-acetyl-alpha-D-muramate	pH 9.0, 37°C, recombinant enzyme MurB/CVs	Verrucomicrobium spinosum
0.47	-	ATP	pH 9.0, 37°C, recombinant enzyme MurB/CVs	Verrucomicrobium spinosum

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytoplasm	-	Verrucomicrobium spinosum	5737	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required, optimal at 10 mM for the UDP-N-acetylmuramate:L-alanine ligase activity of MurB/CVs	Verrucomicrobium spinosum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine	Verrucomicrobium spinosum	-	ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine	-	?
ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine	Verrucomicrobium spinosum DSM 4136	-	ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine	-	?

Organism

Organism	UniProt	Comment	Textmining
Verrucomicrobium spinosum	-	-	-
Verrucomicrobium spinosum DSM 4136	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
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)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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	-	?
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	-	?
ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine	-	Verrucomicrobium spinosum	ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine	-	?
ATP + UDP-N-acetyl-alpha-D-muramate + L-alanine	-	Verrucomicrobium spinosum DSM 4136	ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanine	-	?
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	-	?
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	-	?
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	?	-	?
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

Subunits	Comment	Organism
More	enzyme domain organization, overview	Verrucomicrobium spinosum

Synonyms

Synonyms	Comment	Organism
MurB/CVs	-	Verrucomicrobium spinosum
UDP-N-acetylenolpyruvoylglucosamine reductase/UDP-N-acetylmuramate: L-alanine ligase	-	Verrucomicrobium spinosum
UDP-N-acetylmuramate:L-alanine ligase	-	Verrucomicrobium spinosum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
44	46	UDP-N-acetylmuramate:L-alanine ligase activity of MurB/CVs	Verrucomicrobium spinosum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
9	-	UDP-N-acetylmuramate:L-alanine ligase activity of MurB/CVs	Verrucomicrobium spinosum

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Verrucomicrobium spinosum

General Information

General Information	Comment	Organism
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
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
physiological function	the fusion gene encoding MurB/CVs is able to complement Escherichia coli murB and murC temperature-sensitive mutants	Verrucomicrobium spinosum