Literature summary for 2.4.2.54 extracted from

Bechard, M.E.; Farahani, P.; Greene, D.; Pham, A.; Orry, A.; Rasche, M.E.
Purification, kinetic characterization, and site-directed mutagenesis of Methanothermobacter thermautotrophicus RFAP synthase produced in Escherichia coli (2019), AIMS Microbiol., 5, 186-204 .

Search for more literature for 2.4.2.54

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Methanothermobacter thermautotrophicus

Protein Variants

Protein Variants	Comment	Organism
D19A	inactive	Methanothermobacter thermautotrophicus
D19N	the mutant enzyme with low activity has its temperature optimum at 60°C	Methanothermobacter thermautotrophicus
R26A	inactive	Methanothermobacter thermautotrophicus
R26K	the mutant enzyme with low activity has its temperature optimum at 50°C	Methanothermobacter thermautotrophicus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
3.9	-	4-Aminobenzoate	wild type enzyme, at pH 6.8 and 50°C	Methanothermobacter thermautotrophicus
4.5	-	4-Aminobenzoate	wild type enzyme, at pH 6.8 and 70°C	Methanothermobacter thermautotrophicus
12	-	4-Aminobenzoate	mutant enzyme D19N, at pH 6.8 and 50°C	Methanothermobacter thermautotrophicus
38	-	4-Aminobenzoate	mutant enzyme R26K, at pH 6.8 and 50°C	Methanothermobacter thermautotrophicus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required for activity, 30 mM used in assay conditions	Methanothermobacter thermautotrophicus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
69000	-	gel filtration	Methanothermobacter thermautotrophicus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
5-phospho-alpha-D-ribose 1-diphosphate + 4-aminobenzoate	Methanothermobacter thermautotrophicus	-	beta-ribofuranosylaminobenzene 5'-phosphate + CO2 + diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanothermobacter thermautotrophicus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
hydroxyapatite column chromatography, Mono Q column chromatography and Superdex 75 gel filtration	Methanothermobacter thermautotrophicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.00013	-	cell-free extract, at pH 6.8 and 70°C	Methanothermobacter thermautotrophicus
0.36	-	after 2800fold purification, at pH 6.8 and 70°C	Methanothermobacter thermautotrophicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5-phospho-alpha-D-ribose 1-diphosphate + 4-aminobenzoate	-	Methanothermobacter thermautotrophicus	beta-ribofuranosylaminobenzene 5'-phosphate + CO2 + diphosphate	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * 38000, SDS-PAGE	Methanothermobacter thermautotrophicus

Synonyms

Synonyms	Comment	Organism
beta-ribofuranosylaminobenzene 5'-phosphate synthase	-	Methanothermobacter thermautotrophicus
MTH0830	-	Methanothermobacter thermautotrophicus
RFAP synthase	-	Methanothermobacter thermautotrophicus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
70	-	-	Methanothermobacter thermautotrophicus

General Information

General Information	Comment	Organism
metabolism	the enzyme catalyzes the first committed step in the biosynthesis of tetrahydromethanopterin	Methanothermobacter thermautotrophicus

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Release 2023.1 (January 2023)