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Literature summary extracted from

  • Kuan, Y.; Thiruvengadam, V.; Lin, J.; Liu, J.; Chen, T.; Wu, H.; Wang, W.; Chen, L.
    Broad-specificity amino acid racemase, a novel non-antibiotic selectable marker for transgenic plants (2018), Plant Biotechnol. Rep., 12, 27-38 .
No PubMed abstract available

Application

EC Number Application Comment Organism
5.1.1.10 diagnostics broad-specificity amino acid racemase is a non-antibiotic selectable marker for transgenic plants and can be used in the development of genetically modified plants without using antibiotic resistance marker genes. Selection with different amino acid enantiomers using Bsar, Bsar-R174A and Bsar-R174K gene as selectable markers, overview Pseudomonas putida

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.1.10 recombinant overexpression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) and complementation of lysine or asparagine auxotroph strains (Escherichia coli lysine auxotrophic strain BCRC 51734 and asparagine auxotrophic strain CGSC 4813) recombinant expression of wild-type and mutant enzymes in Arabidopsis thaliana ecotype Columbia and Oryza sativa cv. Tainung 67 plants using the Agrobacterium tumefaciens strain EHA105 transformation system. The Bsar variant, Bsar-R174K, is useful as a selectable marker gene in Arabidopsis and rice that are susceptible to L-lysine and D-alanine. The transformation of Arabidopsis thalinana with Bsar or Bsar variants based on D-alanine selection reveals that Bsar-R174K has the greatest efficiency (2.40%), superior to kanamycin selection-based transformation (1.10%). L-lysine-based selection exhibits lower efficiency for Bsar-R174K (0.17%). The progenies of selected Bsar-R174K transgenic Arabidopsis thaliana plants reveals normal growth properties. Bsar-R174K mutant transgenic Oryza sativa is obtained on L-lysine medium with an efficiency of 0.9%, and the progenies of the transgenic rice show morphologically normal phenotypes comparable with their wild-type counterparts. RT-PCR expression analysis Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
5.1.1.10 R174A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Pseudomonas putida
5.1.1.10 R174K site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme. Mutant Bsar-R174K shows no deleterious effects on the transgenic plants and broad range amino acid racemase activity, might serve as a good selection marker Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.1.1.10 L-alanine Pseudomonas putida
-
 D-alanine
-
 r
5.1.1.10 L-alanine Pseudomonas putida DSM 84
-
 D-alanine
-
 r
5.1.1.10 L-arginine Pseudomonas putida
-
 D-arginine
-
 r
5.1.1.10 L-arginine Pseudomonas putida DSM 84
-
 D-arginine
-
 r
5.1.1.10 L-asparagine Pseudomonas putida
-
 D-asparagine
-
 r
5.1.1.10 L-lysine Pseudomonas putida
-
 D-lysine
-
 r
5.1.1.10 L-lysine Pseudomonas putida DSM 84
-
 D-lysine
-
 r
5.1.1.10 L-Methionine Pseudomonas putida
-
 D-Methionine
-
 r
5.1.1.10 L-Methionine Pseudomonas putida DSM 84
-
 D-Methionine
-
 r
5.1.1.10 L-serine Pseudomonas putida
-
 D-serine
-
 r
5.1.1.10 L-serine Pseudomonas putida DSM 84
-
 D-serine
-
 r

Organism

EC Number Organism UniProt Comment Textmining
5.1.1.10 Pseudomonas putida
-
-
-
5.1.1.10 Pseudomonas putida DSM 84
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.1.1.10 0.1
-
 purified recombinant mutant R174A, pH 7.0, 37°C, substrate L-serine Pseudomonas putida
5.1.1.10 0.12
-
 purified recombinant mutant R174K, pH 7.0, 37°C, substrate L-alanine Pseudomonas putida
5.1.1.10 0.12
-
 purified recombinant mutant R174K, pH 7.0, 37°C, substrate L-methionine Pseudomonas putida
5.1.1.10 0.15
-
 purified recombinant mutant R174A, pH 7.0, 37°C, substrate L-asparagine Pseudomonas putida
5.1.1.10 0.3
-
 purified recombinant mutant R174K, pH 7.0, 37°C, substrate L-serine Pseudomonas putida
5.1.1.10 0.36
-
 purified recombinant mutant R174A, pH 7.0, 37°C, substrate L-methionine Pseudomonas putida
5.1.1.10 0.62
-
 purified recombinant mutant R174A, pH 7.0, 37°C, substrate L-arginine Pseudomonas putida
5.1.1.10 0.64
-
 purified recombinant mutant R174A, pH 7.0, 37°C, substrate L-lysine Pseudomonas putida
5.1.1.10 0.82
-
 purified recombinant mutant R174K, pH 7.0, 37°C, substrate L-asparagine Pseudomonas putida
5.1.1.10 2.46
-
 purified recombinant mutant R174K, pH 7.0, 37°C, substrate L-lysine Pseudomonas putida
5.1.1.10 3.22
-
 purified recombinant mutant R174K, pH 7.0, 37°C, substrate L-arginine Pseudomonas putida
5.1.1.10 7.5
-
 purified recombinant wild-type enzyme, pH 7.0, 37°C, substrate L-asparagine Pseudomonas putida
5.1.1.10 56
-
 purified recombinant wild-type enzyme, pH 7.0, 37°C, substrate L-alanine Pseudomonas putida
5.1.1.10 209
-
 purified recombinant wild-type enzyme, pH 7.0, 37°C, substrate L-serine Pseudomonas putida
5.1.1.10 256
-
 purified recombinant wild-type enzyme, pH 7.0, 37°C, substrate L-methionine Pseudomonas putida
5.1.1.10 1224
-
 purified recombinant wild-type enzyme, pH 7.0, 37°C, substrate L-arginine Pseudomonas putida
5.1.1.10 1423
-
 purified recombinant wild-type enzyme, pH 7.0, 37°C, substrate L-lysine Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.1.10 L-alanine
-
 Pseudomonas putida D-alanine
-
 r
5.1.1.10 L-alanine
-
 Pseudomonas putida DSM 84 D-alanine
-
 r
5.1.1.10 L-arginine
-
 Pseudomonas putida D-arginine
-
 r
5.1.1.10 L-arginine
-
 Pseudomonas putida DSM 84 D-arginine
-
 r
5.1.1.10 L-asparagine
-
 Pseudomonas putida D-asparagine
-
 r
5.1.1.10 L-lysine
-
 Pseudomonas putida D-lysine
-
 r
5.1.1.10 L-lysine
-
 Pseudomonas putida DSM 84 D-lysine
-
 r
5.1.1.10 L-Methionine
-
 Pseudomonas putida D-Methionine
-
 r
5.1.1.10 L-Methionine
-
 Pseudomonas putida DSM 84 D-Methionine
-
 r
5.1.1.10 L-serine
-
 Pseudomonas putida D-serine
-
 r
5.1.1.10 L-serine
-
 Pseudomonas putida DSM 84 D-serine
-
 r

Synonyms

EC Number Synonyms Comment Organism
5.1.1.10 broad-specificity amino acid racemase
-
 Pseudomonas putida
5.1.1.10 Bsar
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.1.1.10 30
-
 assay at Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.1.10 7
-
 assay at Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
5.1.1.10 pyridoxal 5'-phosphate
-
 Pseudomonas putida