Cloned (Comment) | Organism |
---|---|
gene PtGR, DNa and mino acid seuence determination and analysis, sequence comparisons, recombinant expresssion of His-tagged enzyme in Escherichia coli strain BL21 | Paecilomyces sp. 'thermophila' |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, sitting drop vapour diffusion method, the reservoir solution contains 0.1 M MES buffer, pH 6.5, 0.01 M cobalt (II) chloride, and 1.8 M ammonium sulfate, 20°C, X-ray diffraction structure determination and analysis at 1.75 A resolution | Paecilomyces sp. 'thermophila' |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.1 | - |
glyoxylate | pH 7.5, 50°C, recombinant enzyme, with NADPH | Paecilomyces sp. 'thermophila' |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42000 | - |
- |
Paecilomyces sp. 'thermophila' |
78000 | - |
recombinant enzyme, gel filtration | Paecilomyces sp. 'thermophila' |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
glycolate + NADP+ | Paecilomyces sp. 'thermophila' | - |
glyoxylate + NADPH + H+ | - |
? | |
glycolate + NADP+ | Paecilomyces sp. 'thermophila' J18 | - |
glyoxylate + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paecilomyces sp. 'thermophila' | A0A0H3U0Y7 | - |
- |
Paecilomyces sp. 'thermophila' J18 | A0A0H3U0Y7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme 2.7fold from Escherichia coli strain BL21 by nickel affinity chromatography | Paecilomyces sp. 'thermophila' |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
24.2 | - |
purified recombinant enzyme, pH 7.5, 50°C, with NADH | Paecilomyces sp. 'thermophila' |
70.1 | - |
purified recombinant enzyme, pH 7.5, 50°C, with NADPH | Paecilomyces sp. 'thermophila' |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glycolate + NAD+ | - |
Paecilomyces sp. 'thermophila' | glyoxylate + NADH + H+ | - |
? | |
glycolate + NAD+ | - |
Paecilomyces sp. 'thermophila' J18 | glyoxylate + NADH + H+ | - |
? | |
glycolate + NADP+ | - |
Paecilomyces sp. 'thermophila' | glyoxylate + NADPH + H+ | - |
? | |
glycolate + NADP+ | - |
Paecilomyces sp. 'thermophila' J18 | glyoxylate + NADPH + H+ | - |
? | |
additional information | the enzyme is highly specific for glyoxylate, it shows no detectable activity with 4-methyl-2-oxopentanoate, phenylglyoxylate, pyruvate, oxaloacetate, and alpha-ketoglutarate | Paecilomyces sp. 'thermophila' | ? | - |
? | |
additional information | the enzyme is highly specific for glyoxylate, it shows no detectable activity with 4-methyl-2-oxopentanoate, phenylglyoxylate, pyruvate, oxaloacetate, and alpha-ketoglutarate | Paecilomyces sp. 'thermophila' J18 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 35898, sequence calculation, 2 * 42000, SDS-PAGE, recombinant enzyme | Paecilomyces sp. 'thermophila' |
More | the overall structure of the apo-enzyme monomer adopts the typical D-2-hydroxy-aciddehydrogenase fold with a closed conformation, which comprises two alpha/beta/alpha domains, structure analysis, overview | Paecilomyces sp. 'thermophila' |
Synonyms | Comment | Organism |
---|---|---|
glyoxylate reductase | - |
Paecilomyces sp. 'thermophila' |
PtGR | - |
Paecilomyces sp. 'thermophila' |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Paecilomyces sp. 'thermophila' |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
recombinant enzyme, 1 h, 90% activity remaining | Paecilomyces sp. 'thermophila' |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
at 30°C | Paecilomyces sp. 'thermophila' |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4.5 | 10 | recombinant enzyme, 30°C, 1 h, more than 80% of activity is retained | Paecilomyces sp. 'thermophila' |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme utilize either NADPH or NADH as the coenzyme with glyoxylate, but prefers NADPH rather than NADH as an electron donor. The coenzyme specificity is provided by a cationic cluster consisting of N184, R185, and N186. Cofactor binding structure, overview | Paecilomyces sp. 'thermophila' | |
NAD+ | - |
Paecilomyces sp. 'thermophila' | |
NADH | - |
Paecilomyces sp. 'thermophila' | |
NADP+ | - |
Paecilomyces sp. 'thermophila' | |
NADPH | - |
Paecilomyces sp. 'thermophila' |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Paecilomyces sp. 'thermophila' | sequence calculation | - |
6.33 |
General Information | Comment | Organism |
---|---|---|
evolution | the deduced amino acid sequence of the enzyme from Paecilomyes thermophila has low similarities to the reported glyoxylate reductases | Paecilomyces sp. 'thermophila' |
metabolism | glyoxylate reductase is an important enzyme involved in theglyoxylate metabolism in organism | Paecilomyces sp. 'thermophila' |