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5.1.1.3: glutamate racemase

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

Word Map on EC 5.1.1.3

Reaction

L-glutamate
=
D-glutamate

Synonyms

AAR, BAS0806, BAS4379, BcGR, BsGR, BsRacE, CBL/ALR, cystathionine beta-lyase, D-glutamate racemase, DapF, FnGR, GBAA_0847, GBAA_4717, GLR, GluR, glutamate racemase, glutamic acid racemases, GRL, HpMurI, MetC, More, MurI, RACE, RacE1, RacE2, Racemase, glutamate, Rv1338, TmCBL, wMelCBL

ECTree

     5 Isomerases
         5.1 Racemases and epimerases
             5.1.1 Acting on amino acids and derivatives
                5.1.1.3 glutamate racemase

Purification

Purification on EC 5.1.1.3 - glutamate racemase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation
-
ammonium sulfate precipitation, Co2+-affinity column chromatography, and anion-exchange chromatography
-
by using nickel-chelate chromatography
by using nickel-nitrilotriacetic acid affinity chromatography and gel filtration
-
by using of nickel ion affinity chromatography at 4°C
enzyme is purified by using standard chromatographic methods
gene murI, recombinant expression of His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
protein extract is loaded on a HiTrap affinity column charged with Co2+, the His-tags are not removed as the enzyme activities with and without these tags are not significantly different
-
protein extract is purified by using a nickel-chelated Hi-trap chelating column, Hi-trap sp-FF cation-exchange column and by by gel-filtration on a Superdex-200 prep-grade column
RacE1 is expressed as in Escherichia coli with an amino-terminal hexahistidine fusion peptide to facilitate purification via nickel-chelate affinity chromatography
Q81LA8, Q81UL8
recombinant enzyme 9.16fold from Escherichia coli strain BL21(DE3)/pET20b-murI by L-Glu affinity and anion exchange chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage with the PreScission protease, dialysis and another step of nickel affinity chromatography, to homogeneity
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag removal by TEV protease, and gel filtration, to over 95% purity