Cloned (Comment) | Organism |
---|---|
gene clo glxI, cloning in Escherichia coli strain DH5alpha, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Clostridium acetobutylicum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His6-tagged enzyme with bound Zn2+ or Ni2+, sitting drop vapor diffusion, in 20% v/v isopropyl alcohol and 30% w/v PEG3350 in 0.1 M HEPES buffer, pH 7.5, X-ray diffraction structure determination and analysis at 2.45 A and 2.06 A resolution, respectively. The Ni2+-bound non-His-tagged CLO GlxI structure is solved by molecular replacement using the Zn2+-bound structure as search model | Clostridium acetobutylicum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Cd2+ | activates 5% compared to Ni2+ | Clostridium acetobutylicum | |
Co2+ | activates 40% compared to Ni2+ | Clostridium acetobutylicum | |
Mn2+ | activates 5% compared to Ni2+ | Clostridium acetobutylicum | |
additional information | no activation by Zn2+, poor activity with Ca2+ and Mg2+, the active site geometry of the Ni2/Co2-activated enzyme forms an octahedral coordination with one metal atom, two water molecules, and four metal-binding ligands, while the inactive Zn2-bound enzyme form possesses a trigonal bipyramidal geometry with only one water molecule liganded to the metal center | Clostridium acetobutylicum | |
Ni2+ | required, best divalent metal ion | Clostridium acetobutylicum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
15818 | - |
2 * 15818, His6-tagged enzyme, sequence calculation, the enzyme forms two active sites, each within single subunits | Clostridium acetobutylicum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium acetobutylicum | Q97H22 | gene clo glxI | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage og the His-tag by thrombin, and elimination of the tag by benzamidine affinity chromatography | Clostridium acetobutylicum |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 15818, His6-tagged enzyme, sequence calculation, the enzyme forms two active sites, each within single subunits | Clostridium acetobutylicum |
Synonyms | Comment | Organism |
---|---|---|
CLO GlxI | - |
Clostridium acetobutylicum |
Glx1 | - |
Clostridium acetobutylicum |
glyoxalase I | - |
Clostridium acetobutylicum |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Clostridium acetobutylicum | His6-tagged enzyme, sequence calculation | - |
6.5 |
General Information | Comment | Organism |
---|---|---|
metabolism | the glyoxalase system catalyzes the conversion of toxic, metabolically produced 2-oxoaldehydes, such as methylglyoxal, into their corresponding nontoxic 2-hydroxycarboxylic acids, leading to detoxification of these cellular metabolites | Clostridium acetobutylicum |
physiological function | the glyoxalase system catalyzes the conversion of toxic, metabolically produced 2-oxoaldehydes, such as methylglyoxal, into their corresponding nontoxic 2-hydroxycarboxylic acids, leading to detoxification of these cellular metabolites | Clostridium acetobutylicum |