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Literature summary for 1.3.5.2 extracted from

  • Marcinkeviciene, J.; Jiang, W.; Locke, G.; Kopcho, L.M.; Rogers, M.J.; Copeland, R.A.
    A second dihydroorotate dehydrogenase (Type A) of the human pathogen Enterococcus faecalis: expression, purification, and steady-state kinetic mechanism (2000), Arch. Biochem. Biophys., 377, 178-186.
    View publication on PubMed

Application

Application Comment Organism
medicine inhibition of pyrimidine biosynthesis by targeting enzyme activity, mechanism for antimicrobial intervention Enterococcus faecalis

Cloned(Commentary)

Cloned (Comment) Organism
-
Enterococcus faecalis
expression in Escherichia coli BL21(DE3) TAP330, a pyrD(-) minus derivative Enterococcus faecalis

Inhibitors

Inhibitors Comment Organism Structure
Barbituric acid competitive to dihydroorotate, dead-end inhibitor Enterococcus faecalis
H2O2 byproduct, tends to inactivate enzyme Enterococcus faecalis
Orotate
-
Enterococcus faecalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.115
-
S-dihydroorotate
-
Enterococcus faecalis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36000
-
2 * 36000, SDS-PAGE Enterococcus faecalis
80000
-
gel filtration Enterococcus faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-dihydroorotate + acceptor Enterococcus faecalis reduction of a number of substrates: fumatate, coenzyme ubiquinone-0 and menadione orotate + reduced acceptor
-
?

Organism

Organism UniProt Comment Textmining
Enterococcus faecalis
-
-
-
Enterococcus faecalis
-
family 1 enzyme A
-

Purification (Commentary)

Purification (Comment) Organism
streptomycin sulfate precipitation, anion-exchange, gel filtration Enterococcus faecalis

Reaction

Reaction Comment Organism Reaction ID
(S)-dihydroorotate + a quinone = orotate + a quinol 68% identity with type A enzyme of Lactococcus lactis, two-site non-classical ping-pong kinetic mechanism Enterococcus faecalis

Storage Stability

Storage Stability Organism
-20°C, 50% glycerol, 1 mM DTT, 0.01 mM FMN, no loss of activity during time of studies Enterococcus faecalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-dihydroorotate + acceptor fumarate and menadione as electron acceptors Enterococcus faecalis orotate + reduced acceptor
-
?
(S)-dihydroorotate + acceptor reduction of a number of substrates: fumatate, coenzyme ubiquinone-0 and menadione Enterococcus faecalis orotate + reduced acceptor
-
?
dihydroorotate + acceptor acceptor: menadione Enterococcus faecalis orotate + reduced acceptor
-
?
dihydroorotate + acceptor acceptor: fumarate, coenzyme Q10 Enterococcus faecalis orotate + reduced acceptor
-
?
dihydroorotate + acceptor mechanism and pH-dependence of reaction Enterococcus faecalis orotate + reduced acceptor
-
?

Subunits

Subunits Comment Organism
dimer 2 * 36000, SDS-PAGE Enterococcus faecalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
-
Enterococcus faecalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Enterococcus faecalis

Cofactor

Cofactor Comment Organism Structure
FMN
-
Enterococcus faecalis
FMN flavoprotein Enterococcus faecalis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.003
-
Barbituric acid
-
Enterococcus faecalis
0.0388
-
Orotate
-
Enterococcus faecalis