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Literature summary 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.
    View publication on PubMed

General Stability

EC Number General Stability Organism
2.4.2.54 2 mM dithiothreitol and 10% (v/v) glycerol stabilize the enzyme activity during long-term storage and subsequent purification steps Methanosarcina thermophila

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.2.54 4-(methylamino)benzoic acid 5 mM, complete loss of activity Methanosarcina thermophila
2.4.2.54 4-aminothiobenzoate 5 mM, 24% loss of activity Methanosarcina thermophila
2.4.2.54 4-Bromobenzoate 5 mM, 60% loss of activity Methanosarcina thermophila
2.4.2.54 4-hydroxybenzoate 5 mM, 88% loss of activity Methanosarcina thermophila
2.4.2.54 4-Methoxybenzoate 5 mM, 55% loss of activity Methanosarcina thermophila
2.4.2.54 benzoate 5 mM, 31% loss of activity Methanosarcina thermophila
2.4.2.54 diphosphate 5 mM, 84% inhibition Methanosarcina thermophila
2.4.2.54 additional information no inhibition by carbonyl reagents, 4-aminobenzoic acid methyl ester, aniline, 4-aminobenzamide Methanosarcina thermophila
2.4.2.54 phosphate 10 mM, 37% inhibition Methanosarcina thermophila
2.4.2.54 pyridoxal 5'-phosphate 2.5 mM, 21% inhibition Methanosarcina thermophila
2.4.2.54 ribose 5-phosphate 10 mM, 20% inhibition Methanosarcina thermophila
2.4.2.54 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.54 0.058
-
 4-Aminobenzoate pH 4.8, 50°C Methanosarcina thermophila
2.4.2.54 3.6
-
 5-phospho-alpha-D-ribose 1-diphosphate pH 4.8, 50°C Methanosarcina thermophila

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.54 64000
-
 1 * 64000, SDS-PAGE Methanosarcina thermophila
2.4.2.54 65000
-
 gel filtration Methanosarcina thermophila

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.54 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

EC Number Organism UniProt Comment Textmining
2.4.2.54 Methanosarcina thermophila
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
2.4.2.54 relatively stable in air Methanosarcina thermophila

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.54 partial Methanosarcina thermophila

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.2.54 0.03
-
 pH 4.8, 50°C Methanosarcina thermophila

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.54 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
-
 ?
2.4.2.54 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
-
 ?
2.4.2.54 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

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

Synonyms

EC Number Synonyms Comment Organism
2.4.2.54 beta-RFA-P synthase
-
 Methanosarcina thermophila

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.54 50
-
 assay at Methanosarcina thermophila

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.54 4.8
-
-
 Methanosarcina thermophila

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.4.2.54 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
2.4.2.54 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
2.4.2.54 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]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.4.2.54 0.07
-
 4-hydroxybenzoate pH 4.8, 50°C, Ki(slope) Methanosarcina thermophila
2.4.2.54 0.59
-
 4-Bromobenzoate pH 4.8, 50°C, Ki(slope) Methanosarcina thermophila