Literature summary for 2.4.2.54 extracted from

Rasche, M.E.; White, R.H.
Mechanism for the enzymatic formation of 4-(beta-D-ribofuranosyl)aminobenzene 5'-phosphate during the biosynthesis of methanopterin (1998), Biochemistry, 37, 11343-1151.

Search for more literature for 2.4.2.54

General Stability

General Stability	Organism
2 mM dithiothreitol and 10% (v/v) glycerol stabilize the enzyme activity during long-term storage and subsequent purification steps	Methanosarcina thermophila

Inhibitors

Inhibitors	Comment	Organism	Structure
4-(methylamino)benzoic acid	5 mM, complete loss of activity	Methanosarcina thermophila
4-aminothiobenzoate	5 mM, 24% loss of activity	Methanosarcina thermophila
4-Bromobenzoate	5 mM, 60% loss of activity	Methanosarcina thermophila
4-hydroxybenzoate	5 mM, 88% loss of activity	Methanosarcina thermophila
4-Methoxybenzoate	5 mM, 55% loss of activity	Methanosarcina thermophila
benzoate	5 mM, 31% loss of activity	Methanosarcina thermophila
diphosphate	5 mM, 84% inhibition	Methanosarcina thermophila
additional information	no inhibition by carbonyl reagents, 4-aminobenzoic acid methyl ester, aniline, 4-aminobenzamide	Methanosarcina thermophila
phosphate	10 mM, 37% inhibition	Methanosarcina thermophila
pyridoxal 5'-phosphate	2.5 mM, 21% inhibition	Methanosarcina thermophila
ribose 5-phosphate	10 mM, 20% inhibition	Methanosarcina thermophila
Sodium cyanoborohydride	160 mM, complete inhibition, decreased production of 4-(beta-D-ribofuranosyl)aminobenzene 5'-phosphate and the accumulation of a reduced form of the proposed cyclohexadienimine reaction intermediates	Methanosarcina thermophila

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.058	-	4-Aminobenzoate	pH 4.8, 50°C	Methanosarcina thermophila
3.6	-	5-phospho-alpha-D-ribose 1-diphosphate	pH 4.8, 50°C	Methanosarcina thermophila

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	reaction is dependent on Mg2+ or Mn2+, optimal concentration: 10 mM	Methanosarcina thermophila
Mn2+	reaction is dependent on Mg2+ or Mn2+	Methanosarcina thermophila
additional information	requires no dissociable trace metal ions such as iron or cobalt ion for activity	Methanosarcina thermophila

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
64000	-	1 * 64000, SDS-PAGE	Methanosarcina thermophila
65000	-	gel filtration	Methanosarcina thermophila

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	Methanosarcina thermophila	the enzyme is involved in biosynthesis of tetrahydromethanopterin, a folate analog involved in the C1 metabolism of methanogenic archaea, sulfate-reducing archaea, and methylotrophic bacteria	4-(beta-D-ribofuranosyl)aminobenzene 5'-phosphate + CO2 + diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanosarcina thermophila	-	-	-

Oxidation Stability

Oxidation Stability	Organism
relatively stable in air	Methanosarcina thermophila

Purification (Commentary)

Purification (Comment)	Organism
partial	Methanosarcina thermophila

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.03	-	pH 4.8, 50°C	Methanosarcina thermophila

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5-phospho-alpha-D-ribose 1-diphosphate + 4-aminobenzoate	the enzyme is involved in biosynthesis of tetrahydromethanopterin, a folate analog involved in the C1 metabolism of methanogenic archaea, sulfate-reducing archaea, and methylotrophic bacteria	Methanosarcina thermophila	4-(beta-D-ribofuranosyl)aminobenzene 5'-phosphate + CO2 + diphosphate	-	?
5-phospho-alpha-D-ribose 1-diphosphate + 4-aminobenzoate	sequential pattern of substrate binding. An SN1-like reaction produces oxonium ion character at C-1 of 5-phospho-alpha-D-ribose 1-diphosphate which undergoes an ipso electrophilic aromatic substitution reaction at the carboxylic acid-bound carbon of 4-aminobenzoate. Decarboxylation of the resulting cyclohexadienimine intermediate leads to the formation of 4-(beta-D-ribofuranosyl)aminobenzene 5'-phosphate	Methanosarcina thermophila	4-(beta-D-ribofuranosyl)aminobenzene 5'-phosphate + CO2 + diphosphate	-	?
5-phospho-alpha-D-ribose 1-diphosphate + 4-hydroxybenzoate	sequential pattern of substrate binding	Methanosarcina thermophila	4-(beta-D-ribofuranosyl)hydroxybenzene 5'-phosphate + CO2 + diphosphate	-	?

Subunits

Subunits	Comment	Organism
monomer	1 * 64000, SDS-PAGE	Methanosarcina thermophila

Synonyms

Synonyms	Comment	Organism
beta-RFA-P synthase	-	Methanosarcina thermophila

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	assay at	Methanosarcina thermophila

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4.8	-	-	Methanosarcina thermophila

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
4	-	the activity of the enzyme pretreated for 30 min at pH 4.0 is 48% of the activity for enzyme treated at either pH 4.8 or 5.9	Methanosarcina thermophila
4.8	-	the activity of the enzyme pretreated for 30 min at pH 4.0 is 48% of the activity for enzyme treated at either pH 4.8 or 5.9	Methanosarcina thermophila
5.9	-	the activity of the enzyme pretreated for 30 min at pH 4.0 is 48% of the activity for enzyme treated at either pH 4.8 or 5.9	Methanosarcina thermophila

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.07	-	4-hydroxybenzoate	pH 4.8, 50°C, Ki(slope)	Methanosarcina thermophila
0.59	-	4-Bromobenzoate	pH 4.8, 50°C, Ki(slope)	Methanosarcina thermophila

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Release 2023.1 (January 2023)