Literature summary for 4.1.99.12 extracted from

Fischer, M.; Romisch, W.; Schiffmann, S.; Kelly, M.; Oschkinat, H.; Steinbacher, S.; Huber, R.; Eisenreich, W.; Richter, G.; Bacher, A.
Biosynthesis of riboflavin in archaea studies on the mechanism of 3,4-dihydroxy-2-butanone-4-phosphate synthase of Methanococcus jannaschii (2002), J. Biol. Chem., 277, 41410-41416.

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Cloned(Commentary)

Cloned (Comment)	Organism
DNA and amino acid sequence determination and analysis, expression of wild-type and mutant enzymes in Escherichia coli strains XL-1 Blue and M15	Methanocaldococcus jannaschii

Protein Variants

Protein Variants	Comment	Organism
D21E	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
D21N	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
D30E	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
E185D	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
E26D	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
E26Q	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
E28D	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii
H164N	site-directed mutagenesis, inactive mutant	Methanocaldococcus jannaschii

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Methanocaldococcus jannaschii

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
25799	-	2 * 25799, recombinant enzyme, mass spectrometry and SDS-PAGE	Methanocaldococcus jannaschii
51600	-	recombinant enzyme, analytical ultrafiltration	Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-ribulose 5-phosphate	Methanocaldococcus jannaschii	-	formate + L-3,4-dihydroxy-2-butanone-4-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanocaldococcus jannaschii	-	orf MJ0055	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli by hydrophobic interaction and hydroxyapatite chromatography, and ultrafiltration	Methanocaldococcus jannaschii

Reaction

Reaction	Comment	Organism	Reaction ID
D-ribulose 5-phosphate = formate + L-3,4-dihydroxybutan-2-one 4-phosphate	reaction mechanism involving intramolecular skeletal rearrangement, a cluster of charged amino acid residues comprising Arg25, Glu26 and Glu28, Asp21 and Asp30 is essential for catalytic activity, as well as His164 and Glu185	Methanocaldococcus jannaschii	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.174	-	purified enzyme	Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-ribulose 5-phosphate	-	Methanocaldococcus jannaschii	formate + L-3,4-dihydroxy-2-butanone-4-phosphate	-	?
D-ribulose 5-phosphate	the reaction involves an intramolecular skeletal rearrangement, NMR study	Methanocaldococcus jannaschii	formate + L-3,4-dihydroxy-2-butanone-4-phosphate	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 25799, recombinant enzyme, mass spectrometry and SDS-PAGE	Methanocaldococcus jannaschii

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	7.7	assay at	Methanocaldococcus jannaschii

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Release 2023.1 (January 2023)