Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.2.1.6 extracted from

  • Lee, S.C.; Jung, I.P.; Baig, I.A.; Chien, P.N.; La, I.J.; Yoon, M.Y.
    Mutational analysis of critical residues of FAD-independent catabolic acetolactate synthase from Enterococcus faecalis V583 (2015), Int. J. Biol. Macromol., 72, 104-109.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information the enzyme is a FAD-independent catabolic enzyme form Enterococcus faecalis

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Enterococcus faecalis
gene als, genetic location within the butanediol operon, sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Enterococcus faecalis

Protein Variants

Protein Variants Comment Organism
H111F site-directed mutagenesis, the mutant enzyme exhibits about 50fold lower specific activity with significant reduction in kcat compared to the wild-type enzyme Enterococcus faecalis
H111F the mutant displays 26fold increase in Km value compared to the wild type enzyme Enterococcus faecalis
H111R site-directed mutagenesis, the mutant enzyme exhibits increasedspecific activity and kcat compared to the wild-type enzyme Enterococcus faecalis
H111R the mutant displays 17fold increase in Km value compared to the wild type enzyme Enterococcus faecalis
Q112E site-directed mutagenesis, the mutant enzyme exhibits about 50fold lower specific activity with significant reduction in kcat compared to the wild-type enzyme Enterococcus faecalis
Q112E the mutant exhibits significantly lower specific activity with 70fold higher Ks for thiamine diphosphate compared to the wild type enzyme Enterococcus faecalis
Q112N site-directed mutagenesis, the mutant enzyme exhibits about 50fold lower specific activity with significant reduction in kcat compared to the wild-type enzyme Enterococcus faecalis
Q112N the mutant exhibits significantly lower specific activity with 15fold higher Ks for thiamine diphosphate compared to the wild type enzyme Enterococcus faecalis
Q112V site-directed mutagenesis, the mutant enzyme exhibits about 50fold lower specific activity with significant reduction in kcat compared to the wild-type enzyme Enterococcus faecalis
Q112V the mutant exhibits significantly lower specific activity with 10fold higher Ks for thiamine diphosphate compared to the wild type enzyme Enterococcus faecalis
Q411E site-directed mutagenesis, the mutant enzyme exhibits increased specific activity and kcat compared to the wild-type enzyme Enterococcus faecalis
Q411E the mutant shows a 10fold rise in Km and a 20fold increase in Ks for thiamine diphosphate compared to the wild type enzyme Enterococcus faecalis
Q411N site-directed mutagenesis, the mutant enzyme has a 3fold increased Km compared to the wild-type enzyme Enterococcus faecalis
Q411N the mutant exhibits increased specific activity and kcat compared to the wild type enzyme Enterococcus faecalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information binding kinetics of Mg2+ and thiamine diphosphate with wild-type enzyme and mutant enzymes, overview Enterococcus faecalis
1.37
-
pyruvate pH 7.5, 37°C, recombinant wild-type enzyme Enterococcus faecalis
1.37
-
pyruvate wild type enzyme, at pH 6.8 and 37°C Enterococcus faecalis
4.34
-
pyruvate pH 7.5, 37°C, recombinant mutant Q411N Enterococcus faecalis
4.34
-
pyruvate mutant enzyme Q411N, at pH 6.8 and 37°C Enterococcus faecalis
14.68
-
pyruvate pH 7.5, 37°C, recombinant mutant Q112N Enterococcus faecalis
14.68
-
pyruvate mutant enzyme Q112N, at pH 6.8 and 37°C Enterococcus faecalis
16.43
-
pyruvate pH 7.5, 37°C, recombinant mutant Q411E Enterococcus faecalis
16.43
-
pyruvate mutant enzyme Q411E, at pH 6.8 and 37°C Enterococcus faecalis
19.78
-
pyruvate pH 7.5, 37°C, recombinant mutant Q112E Enterococcus faecalis
19.78
-
pyruvate mutant enzyme Q112E, at pH 6.8 and 37°C Enterococcus faecalis
24.6
-
pyruvate pH 7.5, 37°C, recombinant mutant H111R Enterococcus faecalis
24.6
-
pyruvate mutant enzyme H111R, at pH 6.8 and 37°C Enterococcus faecalis
31.2
-
pyruvate pH 7.5, 37°C, recombinant mutant Q112V Enterococcus faecalis
31.2
-
pyruvate mutant enzyme Q112V, at pH 6.8 and 37°C Enterococcus faecalis
36.6
-
pyruvate pH 7.5, 37°C, recombinant mutant H111F Enterococcus faecalis
36.6
-
pyruvate mutant enzyme H111F, at pH 6.8 and 37°C Enterococcus faecalis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ divalent metal ion Mg2+ i required for catalytic activity. In catalysis, a divalent metal ion Mg2+ serves to anchor the diphosphate moiety of thiamine diphosphate at the active site of the catabolic enzyme Enterococcus faecalis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
60000
-
x * 60000, SDS-PAGE Enterococcus faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 pyruvate Enterococcus faecalis
-
2-acetolactate + CO2
-
?

Organism

Organism UniProt Comment Textmining
Enterococcus faecalis Q836A5
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni+-chelating column chromatography Enterococcus faecalis
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) Enterococcus faecalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 pyruvate
-
Enterococcus faecalis 2-acetolactate + CO2
-
?

Subunits

Subunits Comment Organism
? x * 60000, SDS-PAGE Enterococcus faecalis
More the catabolic enzyme form lacks a regulatory subunit Enterococcus faecalis

Synonyms

Synonyms Comment Organism
ALS
-
Enterococcus faecalis
CalS
-
Enterococcus faecalis
catabolic acetolactate synthase
-
Enterococcus faecalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Enterococcus faecalis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.08
-
pyruvate pH 7.5, 37°C, recombinant mutant Q112E Enterococcus faecalis
0.08
-
pyruvate pH 7.5, 37°C, recombinant mutant Q112V Enterococcus faecalis
0.08
-
pyruvate mutant enzyme Q112E, at pH 6.8 and 37°C Enterococcus faecalis
0.08
-
pyruvate mutant enzyme Q112V, at pH 6.8 and 37°C Enterococcus faecalis
0.22
-
pyruvate pH 7.5, 37°C, recombinant mutant Q112N Enterococcus faecalis
0.22
-
pyruvate mutant enzyme Q112N, at pH 6.8 and 37°C Enterococcus faecalis
0.49
-
pyruvate pH 7.5, 37°C, recombinant mutant H111F Enterococcus faecalis
0.49
-
pyruvate mutant enzyme H111F, at pH 6.8 and 37°C Enterococcus faecalis
2.88
-
pyruvate pH 7.5, 37°C, recombinant mutant Q411E Enterococcus faecalis
2.88
-
pyruvate mutant enzyme Q411E, at pH 6.8 and 37°C Enterococcus faecalis
5.28
-
pyruvate pH 7.5, 37°C, recombinant wild-type enzyme Enterococcus faecalis
5.28
-
pyruvate wild type enzyme, at pH 6.8 and 37°C Enterococcus faecalis
15.9
-
pyruvate pH 7.5, 37°C, recombinant mutant Q411N Enterococcus faecalis
15.9
-
pyruvate mutant enzyme Q411N, at pH 6.8 and 37°C Enterococcus faecalis
28.6
-
pyruvate pH 7.5, 37°C, recombinant mutant H111R Enterococcus faecalis
28.6
-
pyruvate mutant enzyme H111R, at pH 6.8 and 37°C Enterococcus faecalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
Enterococcus faecalis

Cofactor

Cofactor Comment Organism Structure
additional information FAD-independent enzyme Enterococcus faecalis
thiamine diphosphate
-
Enterococcus faecalis
thiamine diphosphate dependent on, located centrally in the active site of cALS with a unique V-conformation at the dimer interface to play a central role of intramolecular protontransfer in the catalytic cycle. In catalysis, a divalent metal ion Mg2+ serves to anchor the diphosphate moiety of thiamine diphosphate at the active site of the catalbolic enzyme Enterococcus faecalis

General Information

General Information Comment Organism
additional information the catabolic enzyme form lacks a regulatory subunit. Residue His 111, which is widely present as phenylalanine in many other ThDP-dependent enzymes, plays a crucial role in substrate binding, importance of residues H111, Q112, and Q411 residues for catalysis, Q112 and Q411 might be involved in thiamine diphosphate binding, enzyme structure homology modeling, overview Enterococcus faecalis
physiological function catabolic acetolactate synthase from Enterococcus faecalis is a FAD-independent enzyme, which catalyzes the condensation of two molecules of pyruvate to produce acetolactate Enterococcus faecalis