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Literature summary for 2.7.2.4 extracted from

  • Qi, Q.; Huang, J.; Crowley, J.; Ruschke, L.; Goldman, B.S.; Wen, L.; Rapp, W.D.
    Metabolically engineered soybean seed with enhanced threonine levels: biochemical characterization and seed-specific expression of lysine-insensitive variants of aspartate kinases from the enteric bacterium Xenorhabdus bovienii (2011), Plant Biotechnol. J., 9, 193-204.
    View publication on PubMed
Search for more literature for 2.7.2.4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Xenorhabdus bovienii

Protein Variants

Protein Variants Comment Organism
E257K mutation of the conserved Glu-257 to Lys or the double mutations T359I/E257K render the enzyme insensitive to L-lysine with 86- and 112fold increases in IC50 values, respectively, when compared to the wild-type enzyme. E257K and E257K/T359I alleles exhibit a 1.2- to 1.7fold decrease in Vmax value and 2- to 6fold decrease in kcat/Km value for either substrate compared to wild-type Xenorhabdus bovienii
T359I mutation increases the L-lysine IC50 value by 104fold, but no substantial differences are observed in kinetic parameters except lower Km ATP value compared to the wild-type enzyme. Seed-specific expression of the feedback-resistant mutant T359I or mutant E257K results in increases of free L-threonine levels of up to 100fold in R1 soybean seed when compared to wild-type Xenorhabdus bovienii
T359I/E257K mutation of the conserved Glu-257 to Lys or the double mutations T359I/E257K render the enzyme insensitive to L-lysine with 86- and 112fold increases in IC50 values, respectively, when compared to the wild-type enzyme. E257K and E257K/T359I alleles exhibit a 1.2- to 1.7fold decrease in Vmax value and 2- to 6fold decrease in kcat/Km value for either substrate compared to wild-type Xenorhabdus bovienii

Inhibitors

Inhibitors Comment Organism Structure
L-lysine
-
 Xenorhabdus bovienii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.62
-
 ATP pH 8.0, 30°C, mutant E257K/T359I Xenorhabdus bovienii
1.82
-
 ATP pH 8.0, 30°C, mutant T359I Xenorhabdus bovienii
1.89
-
 ATP pH 8.0, 30°C, mutant E257K Xenorhabdus bovienii
2.31
-
 L-aspartate pH 8.0, 30°C, mutant T359I Xenorhabdus bovienii
2.56
-
 ATP pH 8.0, 30°C, wild-type Xenorhabdus bovienii
2.73
-
 L-aspartate pH 8.0, 30°C, wild-type Xenorhabdus bovienii
3.06
-
 L-aspartate pH 8.0, 30°C, mutant E257K Xenorhabdus bovienii
3.1
-
 L-aspartate pH 8.0, 30°C, mutant E257K/T359I Xenorhabdus bovienii

Organism

Organism UniProt Comment Textmining
Xenorhabdus bovienii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Xenorhabdus bovienii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-aspartate
-
 Xenorhabdus bovienii ADP + 4-phospho-L-aspartate
-
 ?

Synonyms

Synonyms Comment Organism
XbAK
-
 Xenorhabdus bovienii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Xenorhabdus bovienii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Xenorhabdus bovienii

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.4
-
 pH 8.0, 30°C, wild-type Xenorhabdus bovienii L-lysine
121.5
-
 pH 8.0, 30°C, mutant E257K Xenorhabdus bovienii L-lysine
145.4
-
 pH 8.0, 30°C, mutant T359I Xenorhabdus bovienii L-lysine
157.2
-
 pH 8.0, 30°C, mutant E257K/T359I Xenorhabdus bovienii L-lysine

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
44
-
 L-aspartate pH 8.0, 30°C, mutant E257K Xenorhabdus bovienii
110
-
 ATP pH 8.0, 30°C, mutant E257K Xenorhabdus bovienii
120
-
 L-aspartate pH 8.0, 30°C, mutant E257K/T359I Xenorhabdus bovienii
170
-
 ATP pH 8.0, 30°C, mutant E257K/T359I Xenorhabdus bovienii
270
-
 L-aspartate pH 8.0, 30°C, wild-type Xenorhabdus bovienii
290
-
 L-aspartate pH 8.0, 30°C, mutant T359I Xenorhabdus bovienii
310
-
 ATP pH 8.0, 30°C, wild-type Xenorhabdus bovienii
370
-
 ATP pH 8.0, 30°C, mutant T359I Xenorhabdus bovienii