Literature summary extracted from

Dong, H.; Hu, T.; He, G.; Lu, D.; Qi, J.; Dou, Y.; Long, W.; He, X.; Ju, J.; Su, D.
Structural features and kinetic characterization of alanine racemase from Bacillus pseudofirmus OF4 (2018), Biochem. Biophys. Res. Commun., 497, 139-145 .

Application

EC Number	Application	Comment	Organism
5.1.1.1	drug development	the enzyme is an attractive target for designing antibacterial drugs	Alkalihalophilus pseudofirmus

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
5.1.1.1	gene dadX, recombinant expression of wild-type and mutant enzymes	Alkalihalophilus pseudofirmus

Crystallization (Commentary)

EC Number	Crystallization (Comment)	Organism
5.1.1.1	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

EC Number	Protein Variants	Comment	Organism
5.1.1.1	D318K	site-directed mutagenesis, almost inactive mutant, the mutant shows an altered structure compared to the wild-type enzyme	Alkalihalophilus pseudofirmus
5.1.1.1	D43K	site-directed mutagenesis, the mutant shows an altered structure and reduced activity compared to the wild-type enzyme	Alkalihalophilus pseudofirmus
5.1.1.1	D70K	site-directed mutagenesis, the mutant shows an altered structure and reduced activity compared to the wild-type enzyme	Alkalihalophilus pseudofirmus
5.1.1.1	E134K	site-directed mutagenesis, almost inactive mutant, the mutant shows an altered structure compared to the wild-type enzyme	Alkalihalophilus pseudofirmus
5.1.1.1	E71K	site-directed mutagenesis, almost inactive mutant, the mutant shows an altered structure compared to the wild-type enzyme	Alkalihalophilus pseudofirmus

KM Value [mM]

EC Number	KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
5.1.1.1	additional information	-	additional information	enzyme kinetics analysis	Alkalihalophilus pseudofirmus
5.1.1.1	20.78	-	D-alanine	pH 10.5, 40°C, recombinant wild-type enzyme	Alkalihalophilus pseudofirmus
5.1.1.1	56.17	-	L-alanine	pH 10.5, 40°C, recombinant wild-type enzyme	Alkalihalophilus pseudofirmus

Molecular Weight [Da]

EC Number	Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
5.1.1.1	45000	-	mutant E71K, gel filtration	Alkalihalophilus pseudofirmus
5.1.1.1	48000	-	mutant E134K, gel filtration	Alkalihalophilus pseudofirmus
5.1.1.1	49000	-	mutant D318K, gel filtration	Alkalihalophilus pseudofirmus
5.1.1.1	69000	-	mutant D70K, gel filtration	Alkalihalophilus pseudofirmus
5.1.1.1	76000	-	mutant D43K, gel filtration	Alkalihalophilus pseudofirmus
5.1.1.1	78630	-	recombinant wild-type enzyme, gel filtration	Alkalihalophilus pseudofirmus
5.1.1.1	80640	-	sequence calculation, wild-type enzyme	Alkalihalophilus pseudofirmus

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
5.1.1.1	L-alanine	Alkalihalophilus pseudofirmus	-	D-alanine	-	r
5.1.1.1	L-alanine	Alkalihalophilus pseudofirmus OF4	-	D-alanine	-	r

Organism

EC Number	Organism	UniProt	Comment	Textmining
5.1.1.1	Alkalihalophilus pseudofirmus	B3VI72	-	-
5.1.1.1	Alkalihalophilus pseudofirmus OF4	B3VI72	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
5.1.1.1	recombinant wild-type and mutant enzymes	Alkalihalophilus pseudofirmus

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5.1.1.1	D-alanine	reversible racemization	Alkalihalophilus pseudofirmus	L-alanine	-	r
5.1.1.1	D-alanine	reversible racemization	Alkalihalophilus pseudofirmus OF4	L-alanine	-	r
5.1.1.1	L-alanine	-	Alkalihalophilus pseudofirmus	D-alanine	-	r
5.1.1.1	L-alanine	reversible racemization	Alkalihalophilus pseudofirmus	D-alanine	-	r
5.1.1.1	L-alanine	-	Alkalihalophilus pseudofirmus OF4	D-alanine	-	r
5.1.1.1	L-alanine	reversible racemization	Alkalihalophilus pseudofirmus OF4	D-alanine	-	r

Subunits

EC Number	Subunits	Comment	Organism
5.1.1.1	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

EC Number	Synonyms	Comment	Organism
5.1.1.1	ALR	-	Alkalihalophilus pseudofirmus
5.1.1.1	dadXOF4	-	Alkalihalophilus pseudofirmus

Temperature Optimum [°C]

EC Number	Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
5.1.1.1	40	-	-	Alkalihalophilus pseudofirmus

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5.1.1.1	10.5	-	-	Alkalihalophilus pseudofirmus

Cofactor

EC Number	Cofactor	Comment	Organism	Structure
5.1.1.1	pyridoxal 5'-phosphate	PLP, dependent on	Alkalihalophilus pseudofirmus

General Information

EC Number	General Information	Comment	Organism
5.1.1.1	malfunction	enzyme Alr inhibition is lethal to prokaryotes	Alkalihalophilus pseudofirmus
5.1.1.1	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
5.1.1.1	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

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)