Literature summary for 4.4.1.5 extracted from

Suttisansanee, U.; Lau, K.; Lagishetty, S.; Rao, K.N.; Swaminathan, S.; Sauder, J.M.; Burley, S.K.; Honek, J.F.
Structural variation in bacterial glyoxalase I enzymes: investigation of the metalloenzyme glyoxalase I from Clostridium acetobutylicum (2011), J. Biol. Chem., 286, 38367-38374.

Cloned(Commentary)

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 (Commentary)

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

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

Metals/Ions	Comment	Organism
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 [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

Organism	UniProt	Comment	Textmining
Clostridium acetobutylicum	Q97H22	gene clo glxI	-

Purification (Commentary)

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

Subunits	Comment	Organism
dimer	2 * 15818, His6-tagged enzyme, sequence calculation, the enzyme forms two active sites, each within single subunits	Clostridium acetobutylicum

Synonyms

Synonyms	Comment	Organism
CLO GlxI	-	Clostridium acetobutylicum
Glx1	-	Clostridium acetobutylicum
glyoxalase I	-	Clostridium acetobutylicum

pI Value

Organism	Comment	pI Value Maximum	pI Value
Clostridium acetobutylicum	His6-tagged enzyme, sequence calculation	-	6.5

General Information

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