Literature summary for 4.1.99.12 extracted from

Li, J.; Hua, Z.; Miao, L.; Jian, T.; Wei, Y.; Shasha, Z.; Shaocheng, Z.; Zhen, G.; Hongpeng, Z.; Ailong, H.; Deqiang, W.
The crystal structure and biochemical properties of DHBPS from Streptococcus pneumoniae, a potential anti-infective target for Gram-positive bacteria (2013), Protein Expr. Purif., 91, 161-168.

Search for more literature for 4.1.99.12

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of wild-type andn mutant soluble His-tagged enzymes in Escherichia coli strain BL21(DE3)	Streptococcus pneumoniae

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme, hanging-drop vapor diffusion method, mixing of 0.001 ml of 5 mg/ml protein in 5 mM Tris-HCl, pH 8.0, and 50 mM NaCl, with 0.001 ml of reservoir solution containing ammonium sulfate 2.0 M and 0.1 M BisTris pH7.02, X-ray diffraction structure determination and analysis at 2.0 A resolution	Streptococcus pneumoniae

Protein Variants

Protein Variants	Comment	Organism
D32A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae
E163A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae
E30A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae
H125A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae
H142A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae
T143A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae
T96A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Streptococcus pneumoniae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.181	-	D-ribulose 5-phosphate	pH 7.5, 37°C	Streptococcus pneumoniae

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Co2+	activates	Streptococcus pneumoniae
Fe3+	the enzyme shows activity in the presence of the trivalent metal ion, Fe3+	Streptococcus pneumoniae
Mg2+	required, best physiological metal activator, the divalent metal ion, Mg2+ plays a crucial role for acid/base catalysis	Streptococcus pneumoniae
Mn2+	activates	Streptococcus pneumoniae
additional information	no activity with Cu2+	Streptococcus pneumoniae
Ni2+	activates	Streptococcus pneumoniae
Pb2+	activates	Streptococcus pneumoniae
Zn2+	activates	Streptococcus pneumoniae

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
35350	-	recombinant enzyme, gel filtration	Streptococcus pneumoniae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-ribulose 5-phosphate	Streptococcus pneumoniae	-	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?
D-ribulose 5-phosphate	Streptococcus pneumoniae TIGR 4	-	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Streptococcus pneumoniae	A0A0H2UN41	gene Sp0176	-
Streptococcus pneumoniae TIGR 4	A0A0H2UN41	gene Sp0176	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant soluble His-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography	Streptococcus pneumoniae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.109	-	substrate D-ribulose 5-phosphate, pH 7.5, 37°C	Streptococcus pneumoniae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-ribulose 5-phosphate	-	Streptococcus pneumoniae	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?
D-ribulose 5-phosphate	-	Streptococcus pneumoniae TIGR 4	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?

Subunits

Subunits	Comment	Organism
dimer	DHBPS exists as a homodimer in solution, and the active sites are located at the dimeric interface formed by charged residues from two monomers	Streptococcus pneumoniae
More	the enzyme comprises one beta-sheet (five-stranded) and eight alpha-helices, adopting a three-layered alpha-beta-alpha sandwich fold	Streptococcus pneumoniae

Synonyms

Synonyms	Comment	Organism
DHBPS	-	Streptococcus pneumoniae
Sp0176	-	Streptococcus pneumoniae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
33	-	-	Streptococcus pneumoniae

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
5	55	activity range, profile overview. Highest activity at 33°C, while at 30°C and 37°C, the activity decreases sharply to approximately 15-30% of the activity at 33°C	Streptococcus pneumoniae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Streptococcus pneumoniae

pH Range

pH Minimum	pH Maximum	Comment	Organism
6	8	narrow optimum, no activity below or above, profile overview	Streptococcus pneumoniae

General Information

General Information	Comment	Organism
metabolism	3,4-dihydroxy-2-butanone-4-phosphate synthase is one of the key enzymes in the biosynthesis of riboflavin	Streptococcus pneumoniae
additional information	the two key enzymes of the riboflavin biosynthetic pathway, DHBPS and GTP cyclohydrolaseII (GCHII), EC 3.5.4.25,are fused together into the bifunctional enzyme encoded by Sp0176 gene of Streptococcus pneumoniae strain TIGR 4, DHBPS active site architecture, overview	Streptococcus pneumoniae

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Release 2023.1 (January 2023)