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Literature summary extracted from

  • Strauch, R.C.; Svedin, E.; Dilkes, B.; Chapple, C.; Li, X.
    Discovery of a novel amino acid racemase through exploration of natural variation in Arabidopsis thaliana (2015), Proc. Natl. Acad. Sci. USA, 112, 11726-11731 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.1.21 gene daar1, genotyping, untargeted metabolite profiling and genome-wide association analysis on 440 natural accessions of Arabidopsis thaliana to establish genetic linkages between metabolites and genes, recombinant expression of His-tagged enzyme DAAR1 in Escherichia coli strain Rosetta 2 pLysS. The affinity tag does not affect enzyme activity or stability Arabidopsis thaliana

Protein Variants

EC Number Protein Variants Comment Organism
5.1.1.21 additional information generation of a T-DNA insertion mutant of gene daar1 leading to highly reduced levels of N-malonyl-D-allo-isoleucine Arabidopsis thaliana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.1.21 17
-
L-isoleucine recombinant His-tagged enzyme, pH 8.0, 22°C Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.1.1.21 L-isoleucine Arabidopsis thaliana
-
D-isoleucine
-
r

Organism

EC Number Organism UniProt Comment Textmining
5.1.1.21 Arabidopsis thaliana Q66GM9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.1.1.21 recombinant His-tagged enzyme DAAR1 from Escherichia coli strain Rosetta 2 pLysS by nickel affinity chromatography Arabidopsis thaliana

Source Tissue

EC Number Source Tissue Comment Organism Textmining
5.1.1.21 leaf
-
Arabidopsis thaliana
-

Storage Stability

EC Number Storage Stability Organism
5.1.1.21 purified recombinant His-tagged enzyme, rapid loss of activity at room temperature and when storing the enzyme at 4°C for overnight Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.1.21 L-isoleucine
-
Arabidopsis thaliana D-isoleucine
-
r
5.1.1.21 L-leucine 1% of the activity with L-isoleucine Arabidopsis thaliana D-leucine
-
r
5.1.1.21 L-valine 5% of the activity with L-isoleucine Arabidopsis thaliana D-valine
-
r
5.1.1.21 additional information the enzyme does not require pyridoxal 5'-phosphate for its activity. Compared with Ile, DAAR1 shows 1 and 5% relative activity toward Leu and Val, respectively. DAAR1 shows no activity toward N-acetyl-L-Ile, a commercially available surrogate for N-malonyl-L-Ile, suggesting that DAAR1 does not use a N-derivatized substrate Arabidopsis thaliana ?
-
?

Synonyms

EC Number Synonyms Comment Organism
5.1.1.21 amino acid racemase
-
Arabidopsis thaliana
5.1.1.21 D-amino acid racemase
-
Arabidopsis thaliana
5.1.1.21 DAAR1
-
Arabidopsis thaliana
5.1.1.21 phenazine biosynthesis PhzC/PhzF family protein UniProt Arabidopsis thaliana
5.1.1.21 PLP-independent racemase
-
Arabidopsis thaliana

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.1.1.21 22
-
assay at room temperature Arabidopsis thaliana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.1.1.21 11.03
-
L-isoleucine recombinant His-tagged enzyme, pH 8.0, 22°C Arabidopsis thaliana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.1.21 8
-
assay at Arabidopsis thaliana

Cofactor

EC Number Cofactor Comment Organism Structure
5.1.1.21 additional information enzyme does not require pyridoxal 5'-phosphate for its activity Arabidopsis thaliana

General Information

EC Number General Information Comment Organism
5.1.1.21 evolution the enzyme is a eukaryotic member of a large and widely conserved phenazine biosynthesis protein PhzF-like protein family and belongs to a D-amino acid racemase gene family. The phenazine biosynthesis-like protein family (PF02567), which is closely related to the DAP epimerase (PF01678), proline racemase (PF05544), and PrpF (PF04303) families Arabidopsis thaliana
5.1.1.21 malfunction a daar1 enzyme knockout mutant shows highly reduced levels of N-malonyl-D-allo-isoleucine Arabidopsis thaliana
5.1.1.21 additional information exploitation of natural metabolic variation by integrating metabolomics with genome-wide association as an approach for functional genomics study of specialized metabolism Arabidopsis thaliana
5.1.1.21 physiological function the amino acid racemase is responsible for the biosynthesis of N-malonyl-D-allo-isoleucine. DAAR1 does not use a N-derivatized substrate and the malonylation of D-amino acids occurs downstream in the metabolic pathway Arabidopsis thaliana

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.1.1.21 0.65
-
L-isoleucine recombinant His-tagged enzyme, pH 8.0, 22°C Arabidopsis thaliana