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

BRENDA support

Literature summary for 5.1.1.1 extracted from

  • Prosser, G.A.; Rodenburg, A.; Khoury, H.; de Chiara, C.; Howell, S.; Snijders, A.P.; de Carvalho, L.P.
    Glutamate racemase is the primary target of beta-chloro-D-alanine in Mycobacterium tuberculosis (2016), Antimicrob. Agents Chemother., 60, 6091-6099 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene alr, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3) Bacillus subtilis
gene alr, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3) Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
beta-Chloro-D-alanine BCDA, its primary target is glutamate racemase, poor activity oagainst alanine racemase activity, potent antituberculosis activity. BCDA does not inhibit the D-alanine pathway in intact cells, consistent with its poor in vitro activity, it is instead an irreversible mechanism-based inactivator of glutamate racemase (MurI), an upstream enzyme in the same early stage of peptidoglycan biosynthesis. Inhibition kinetics, overview. Glutamate racemase (MurI) is a pyridoxal 5'-phosphate-independent racemase and is therefore unable to undergo the same mechanism of inhibition as Alr with BCDA Bacillus subtilis
beta-Chloro-D-alanine BCDA, the compund is a very poor inhibitor of recombinant Mycobacterium tuberculosis Alr, despite having potent antituberculosis activity, its primary target is glutamate racemase, poor activity oagainst alanine racemase activity, potent antituberculosis activity. BCDA does not inhibit the D-alanine pathway in intact cells, consistent with its poor in vitro activity, it is instead an irreversible mechanism-based inactivator of glutamate racemase (MurI), an upstream enzyme in the same early stage of peptidoglycan biosynthesis. Inhibition kinetics, overview. Glutamate racemase (MurI) is a pyridoxal 5'-phosphate-independent racemase and is therefore unable to undergo the same mechanism of inhibition as Alr with BCDA Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7
-
L-alanine pH 7.6, 37°C Mycobacterium tuberculosis
5.9
-
L-alanine pH 7.6, 37°C Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-alanine Bacillus subtilis
-
D-alanine
-
r
L-alanine Mycobacterium tuberculosis
-
D-alanine
-
r
L-alanine Bacillus subtilis 168
-
D-alanine
-
r
L-alanine Mycobacterium tuberculosis ATCC 25618 / H37Rv
-
D-alanine
-
r

Organism

Organism UniProt Comment Textmining
Bacillus subtilis P94556
-
-
Bacillus subtilis 168 P94556
-
-
Mycobacterium tuberculosis P9WPW9
-
-
Mycobacterium tuberculosis ATCC 25618 / H37Rv P9WPW9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage Bacillus subtilis
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-alanine
-
Bacillus subtilis D-alanine
-
r
L-alanine
-
Mycobacterium tuberculosis D-alanine
-
r
L-alanine
-
Bacillus subtilis 168 D-alanine
-
r
L-alanine
-
Mycobacterium tuberculosis ATCC 25618 / H37Rv D-alanine
-
r

Synonyms

Synonyms Comment Organism
MurI
-
Bacillus subtilis
MurI
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Bacillus subtilis
37
-
assay at Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
37
-
L-alanine pH 7.6, 37°C Mycobacterium tuberculosis
1190
-
L-alanine pH 7.6, 37°C Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Bacillus subtilis
7.6
-
assay at Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Mycobacterium tuberculosis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.13
-
beta-Chloro-D-alanine pH 7.6, 37°C Mycobacterium tuberculosis
0.23
-
beta-Chloro-D-alanine pH 7.6, 37°C Bacillus subtilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
10
-
L-alanine pH 7.6, 37°C Mycobacterium tuberculosis
201.7
-
L-alanine pH 7.6, 37°C Bacillus subtilis