Application | Comment | Organism |
---|---|---|
drug development | the enzyme is an attractive target for designing antibacterial drugs | Alkalihalophilus pseudofirmus |
Cloned (Comment) | Organism |
---|---|
gene dadX, recombinant expression of wild-type and mutant enzymes | Alkalihalophilus pseudofirmus |
Crystallization (Comment) | Organism |
---|---|
purified enzyme, hanging drop vapour diffusion, method, X-ray diffraction structure determination and analysis at 1.8 A resolution, molecular replacement method using the crystal structure of native alanine racemase from Bacillus stearothermophilus as search model, PDB ID 1SFT | Alkalihalophilus pseudofirmus |
Protein Variants | Comment | Organism |
---|---|---|
D318K | site-directed mutagenesis, almost inactive mutant, the mutant shows an altered structure compared to the wild-type enzyme | Alkalihalophilus pseudofirmus |
D43K | site-directed mutagenesis, the mutant shows an altered structure and reduced activity compared to the wild-type enzyme | Alkalihalophilus pseudofirmus |
D70K | site-directed mutagenesis, the mutant shows an altered structure and reduced activity compared to the wild-type enzyme | Alkalihalophilus pseudofirmus |
E134K | site-directed mutagenesis, almost inactive mutant, the mutant shows an altered structure compared to the wild-type enzyme | Alkalihalophilus pseudofirmus |
E71K | site-directed mutagenesis, almost inactive mutant, the mutant shows an altered structure compared to the wild-type enzyme | Alkalihalophilus pseudofirmus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | enzyme kinetics analysis | Alkalihalophilus pseudofirmus | |
20.78 | - |
D-alanine | pH 10.5, 40°C, recombinant wild-type enzyme | Alkalihalophilus pseudofirmus | |
56.17 | - |
L-alanine | pH 10.5, 40°C, recombinant wild-type enzyme | Alkalihalophilus pseudofirmus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
mutant E71K, gel filtration | Alkalihalophilus pseudofirmus |
48000 | - |
mutant E134K, gel filtration | Alkalihalophilus pseudofirmus |
49000 | - |
mutant D318K, gel filtration | Alkalihalophilus pseudofirmus |
69000 | - |
mutant D70K, gel filtration | Alkalihalophilus pseudofirmus |
76000 | - |
mutant D43K, gel filtration | Alkalihalophilus pseudofirmus |
78630 | - |
recombinant wild-type enzyme, gel filtration | Alkalihalophilus pseudofirmus |
80640 | - |
sequence calculation, wild-type enzyme | Alkalihalophilus pseudofirmus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-alanine | Alkalihalophilus pseudofirmus | - |
D-alanine | - |
r | |
L-alanine | Alkalihalophilus pseudofirmus OF4 | - |
D-alanine | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alkalihalophilus pseudofirmus | B3VI72 | - |
- |
Alkalihalophilus pseudofirmus OF4 | B3VI72 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes | Alkalihalophilus pseudofirmus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-alanine | reversible racemization | Alkalihalophilus pseudofirmus | L-alanine | - |
r | |
D-alanine | reversible racemization | Alkalihalophilus pseudofirmus OF4 | L-alanine | - |
r | |
L-alanine | - |
Alkalihalophilus pseudofirmus | D-alanine | - |
r | |
L-alanine | reversible racemization | Alkalihalophilus pseudofirmus | D-alanine | - |
r | |
L-alanine | - |
Alkalihalophilus pseudofirmus OF4 | D-alanine | - |
r | |
L-alanine | reversible racemization | Alkalihalophilus pseudofirmus OF4 | D-alanine | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | enzyme molecular structure analysis, the tertiary structure of DadXOF4 is a homodimer comprised of two monomers that interact in a head-to-tail manner. Each monomer is composed of two domains, an eight-stranded alpha/beta-barrel at the N-terminus and a C-terminal domain essentially composed of beta-strands. The dimer interface of DadXOF4 is formed by five pairs of salt bridges, i.e. Asp43-Lys360', Asp70-Lys2', Glu71-Arg361', Glu134-Arg259', and Asp318-Lys41' | Alkalihalophilus pseudofirmus |
Synonyms | Comment | Organism |
---|---|---|
ALR | - |
Alkalihalophilus pseudofirmus |
dadXOF4 | - |
Alkalihalophilus pseudofirmus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Alkalihalophilus pseudofirmus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10.5 | - |
- |
Alkalihalophilus pseudofirmus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | PLP, dependent on | Alkalihalophilus pseudofirmus |
General Information | Comment | Organism |
---|---|---|
malfunction | enzyme Alr inhibition is lethal to prokaryotes | Alkalihalophilus pseudofirmus |
additional information | enzyme molecular structure analysis. The conserved residues at the substrate entryway and the salt bridge at the dimer interface are critical for enzyme activity | Alkalihalophilus pseudofirmus |
physiological function | alanine racemase (Alr) is a pyridoxal 5'-phosphate-dependent (PLP) enzyme that catalyzes a reversible racemization between the enantiomers of alanine. D-Alanine is an indispensable constituent in the biosynthesis of bacterial cell-wall peptidoglycan | Alkalihalophilus pseudofirmus |