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5.1.1.1: alanine racemase

This is an abbreviated version!
For detailed information about alanine racemase, go to the full flat file.

Word Map on EC 5.1.1.1

Reaction

L-alanine
=
D-alanine

Synonyms

1SFT, AAR, alanine racemase, AlaR, ALR, alr-2, ALR1, ALR2, Alr2 racemase, AlrA, AlrAba, AlrBax, AlrMtb, alrTt, ARL, BA0252, BAS0238, CBL/ALR, CdAlr, cystathionine beta-lyase, D-alanine racemase, DadB, DadX, dadXOF4, dal1, EcAlr, EcCBL, EfAlaR, L-Alanine racemase, L-Alanine:D-alanine racemase, MBalr1, MBAlr2, MetC, More, MurI, OEOE_1641, PDB, Racemase, alanine, tAlaRac, TmCBL, wMelCBL

ECTree

     5 Isomerases
         5.1 Racemases and epimerases
             5.1.1 Acting on amino acids and derivatives
                5.1.1.1 alanine racemase

Cloned

Cloned on EC 5.1.1.1 - alanine racemase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
31-54% sequence homologies with Bacillus subtilis and Salmonella typhimurium dadB and alr enzymes
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amplified by PCR, cloned and overexpressed in Escherichia coli BL21, both as native and His-tagged products
amplified by PCR, cloned and overexpressed with pOPINB in Escherichia coli Rosetta pLysS cells, His-tagged product
Q81VF6
amplified from genomic DNA from Enterococcus faecalis via PCR, integrated into pET22b, transformation of Escherichia coli BL21 (DE3) for overexpression as His-tagged protein
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analyzation of alanine racemase-transcripts (RT-PCR) during growth and sporulation of wild-type shows low transcription level during early and higher during late sporulation states, analyzed expression level (immunoblotting) shows only alanine racemase during late sporulating states, growth rate of alr-gene knockout mutant and wild-type are identical, mutant only produces half as many spores as wild-type, mutant and wild-type show same resistance against heat, lysozyme and organic solvents, germination takes place in mutant at lower levels of L-alanine (a suggested germination activator) than in wild-type (D-alanine ist suggested to inhibit germination when unfavourable growth conditions), conversion of phase-bright spores to phase-dark germinating cells takes already place within mother cells of the alr mutant (phase dark to phase light), not in wild-type cells resulting in non-resistant mutant-germinants and resistant wild-type-spores respectively
Q81VF6
cloning of two independent alanine racemases in Escherichia coli: Alr and DadX
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enzyme expression in Escherichia coli strain BL21(DE3)/pETALR
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enzyme expression in Escherichia coli strain BL21(DE3)/pMB1978
Escherichia coli strain BL21 (DE3) was transformed with pET28a–Alr (1 wild-type and 8 single point mutants)
expressed as recombinant monomeric protein in Escherichia coli
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expressed in Escherichia coli as an N-terminal polyhistidine fusion
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expressed in Escherichia coli BL21 cells
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) pLysS cells
expressed in Escherichia coli strain ALA1
expression in Escherichia coli
expression in Escherichia coli as a His-tagged protein
-
expression in Escherichia coli C600
-
expression in Escherichia coli JM109
expression in Escherichia coli SOLR with a plasmid yYOK3
expression in Escherichia coli. Heterologous expression renders Saccharomyces cerevisiae capable of utilization of D-Ala as a nitrogen source but also relieves the yeast from the toxicity of D-Ala
expression of C-terminally His6-tagged selenomethionine-derivatized enzyme in Escherichia coli strain BL21(DE3) in HY medium
expression of soluble His-tagged NusA fusion enzyme in Escherichia coli strain HMS174(lambdaDE3)
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from muscle and hepatopancreas
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gene alr from strain OXA-23, DNA and amino acid sequence determination and analysis, expression of recombinant N-terminally His7-tagged enzyme in Escherichia coli strain BL21(DE3)
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gene alr, expression of the enzyme in Escherichia coli strain BL21
gene alr, quantitative RT-PCR expression analysis
gene alr, recombinant expression in Escherichia coli strain BL21(DE3)
gene alr, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene alr, sequence comparisons, recombinant expression of codon-optimized wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 (pLysS)DE3, recombinant expression of wild-type and mutant enzymes in Escherichia coli alr/dadX double-knockout mutant strain MB2159, the wild-type and K271T mutant CdAlr proteins are able to complement the D-alanine auxotrophy and restore its growth in D-alanine-free medium
gene alr, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-CodonPlusTM(DE3)-RIPL
gene alr, sequence comparisons, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21 Star (DE3)pLysS
gene alr-2, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene alr-2, sequence comparisons, the gene encodes a functional enzyme that can complement the alanine racemase deficiency of Escherichia coli strain MB2795, recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene alr1, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
gene alr2, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
gene alrMB4, expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)
gene dadX, recombinant expression of wild-type and mutant enzymes
Alkalihalophilus pseudofirmus
gene MBalr2, DNA and amino acid sequence determination and analysis, expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
gene metC, phylogenetic analysis, the recombinant His6-tagged enzyme TmCBL is overexpressed in a metC-deficient DELTAmetC Escherichia coli mutant strain and can complement it. The enzyme rescues the alanine racemase knockout, Escherichia coli strain MB2795 (DELTAalrDELTAdadX), as quickly as expressing Escherichia coli ALR itself. Recombinant production of His6-tagged enzyme in a soluble form
gene metC, phylogenetic analysis, the recombinant His6-tagged enzyme wMelCBL is overexpressed in a metC-deficient DELTAmetC Escherichia coli mutant strain and can complement it. The enzyme rescues the alanine racemase knockout, Escherichia coli strain MB2795 (DELTAalrDELTAdadX), as quickly as expressing Escherichia coli ALR itself. Recombinant production of His6-tagged enzyme in a soluble form
gene metC, phylogenetic analysis, the recombinant MBP-tagged enzyme PuCBL is overexpressed in a metC-deficient DELTAmetC Escherichia coli mutant strain and can complement it partially. The enzyme rescues the alanine racemase knockout, Escherichia coli strain MB2795 (DELTAalrDELTAdadX), as quickly as expressing Escherichia coli ALR itself. Attempts to optimize recombinant production of the Pelagibacter ubique enzyme (PuCBL) in a soluble form are unsuccessful
gene metC, phylogenetic analysis. The recombinant enzyme EcCBL is overexpressed in a metC-deficient DELTAmetC Escherichia coli mutant strain and can complement it partially
isolation and sequencing of cDNA clones, nucleotide sequence 1798 bp including 5’- and 3’-nonreading frame, poly-A-tail and open reading frame of 1263 bp, expression in E. coli - transformed with open reading frame sequence in pET32Xa/LIC, produced fusion protein with alanine racemase by Escherichia coli, predicted molecular weight of fusion protein of 62000 Da (SDS-PAGE, immunoblotting)
recombinant expression in D-Alanine auxotrophic Escherichia coli strain MB2159 and DN1686, a D-Ala auxotrophic mutant
recombinant expression in D-Alanine auxotrophic Escherichia coli strains MB2159 and DN1686, a D-Ala auxotrophic mutant
strain has one single enzyme gene, expression in Escherichia coli with His-tag
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subcloned in a D-alanine auxotrophic Escherichia coli strain MB2795 and in Escherichia coli BL21(DE3)