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Literature summary 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 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.54 3.9
-
 4-Aminobenzoate wild type enzyme, at pH 6.8 and 50°C Methanothermobacter thermautotrophicus
2.4.2.54 4.5
-
 4-Aminobenzoate wild type enzyme, at pH 6.8 and 70°C Methanothermobacter thermautotrophicus
2.4.2.54 12
-
 4-Aminobenzoate mutant enzyme D19N, at pH 6.8 and 50°C Methanothermobacter thermautotrophicus
2.4.2.54 38
-
 4-Aminobenzoate mutant enzyme R26K, at pH 6.8 and 50°C Methanothermobacter thermautotrophicus

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.54 69000
-
 gel filtration Methanothermobacter thermautotrophicus

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 Methanothermobacter thermautotrophicus
-
 beta-ribofuranosylaminobenzene 5'-phosphate + CO2 + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.54 Methanothermobacter thermautotrophicus
-
-
-

Purification (Commentary)

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

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 0.00013
-
 cell-free extract, at pH 6.8 and 70°C Methanothermobacter thermautotrophicus
2.4.2.54 0.36
-
 after 2800fold purification, at pH 6.8 and 70°C Methanothermobacter thermautotrophicus

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
-
 Methanothermobacter thermautotrophicus beta-ribofuranosylaminobenzene 5'-phosphate + CO2 + diphosphate
-
 ?

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.54 70
-
-
 Methanothermobacter thermautotrophicus

General Information

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