Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Top10 as His-tag fusion protein | Thermus thermophilus HB27 |
expression of the Myc-His9-tagged enzyme in Escherichia coli | Thermus thermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity at more than 500 mM NaCl, 50% activity in the presence of 900 mM sodium phosphate | Thermus thermophilus HB27 | |
NaCl | complete inhibition at 0.5 M | Thermus thermophilus | |
Sodium phosphate | 50% inhibition at 900 mM | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
NADPH | pH 6.7, 45°C, substrate glyoxylate, purified recombinant enzyme | Thermus thermophilus | |
0.04 | - |
NADPH | with glyoxylate as substrate, pH 6.7, 45°C | Thermus thermophilus HB27 | |
0.076 | - |
NADH | pH 6.7, 45°C, substrate glyoxylate, purified recombinant enzyme | Thermus thermophilus | |
0.076 | - |
NADH | with glyoxylate as substrate, pH 6.7, 45°C | Thermus thermophilus HB27 | |
0.5 | - |
glyoxylate | pH 6.7, 45°C, cofactor NADPH, purified recombinant enzyme | Thermus thermophilus | |
0.5 | - |
glyoxylate | with as NADPH as cofactor, pH 6.7, 45°C | Thermus thermophilus HB27 | |
1.2 | - |
glyoxylate | pH 6.7, 45°C, cofactor NADH, purified recombinant enzyme | Thermus thermophilus | |
1.2 | - |
glyoxylate | with as NADH as cofactor, pH 6.7, 45°C | Thermus thermophilus HB27 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36000 | - |
2 * 36000, SDS-PAGE, native mass by gel filtration (native mass not given in the reference) | Thermus thermophilus HB27 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
glyoxylate + NAD(P)H | Thermus thermophilus | - |
glycolate + NAD(P)+ | - |
? | |
glyoxylate + NAD(P)H | Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | - |
glycolate + NAD(P)+ | - |
? | |
glyoxylate + NADPH + H+ | Thermus thermophilus HB27 | - |
glycolate + NADP+ | - |
ir |
Organic Solvent | Comment | Organism |
---|---|---|
1,5-pentanediol | 25%, half life reduced by approximately 70% | Thermus thermophilus HB27 |
diethylene glycol | half-life in the presence of 25% diethylene glycol significantly longer than that in the absence of an organic solvent | Thermus thermophilus HB27 |
dimethyl sulfoxide | half-life in the presence of 25% dimethyl sulfoxide significantly longer than that in the absence of an organic solvent | Thermus thermophilus HB27 |
Ethanol | 25%, half life reduced by approximately 70% | Thermus thermophilus HB27 |
Glycerol | 25%, unstable | Thermus thermophilus HB27 |
n-decane | 25%, unstable | Thermus thermophilus HB27 |
n-hexane | 25%, unstable | Thermus thermophilus HB27 |
n-octane | 25%, unstable | Thermus thermophilus HB27 |
p-xylene | 25%, unstable | Thermus thermophilus HB27 |
toluene | 25%, unstable | Thermus thermophilus HB27 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Thermus thermophilus HB27 | - |
NBRC code 101085 | - |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His9-tagged enzyme from Escherichia coli by nickel affinity chromatography to homogeneity | Thermus thermophilus |
recombinant protein from Escherichia coli using His-tag | Thermus thermophilus HB27 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glyoxylate + NAD(P)H | - |
Thermus thermophilus | glycolate + NAD(P)+ | - |
? | |
glyoxylate + NAD(P)H | - |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | glycolate + NAD(P)+ | - |
? | |
glyoxylate + NADH + H+ | - |
Thermus thermophilus | glycolate + NAD+ | - |
r | |
glyoxylate + NADH + H+ | - |
Thermus thermophilus HB27 | glycolate + NAD+ | - |
ir | |
glyoxylate + NADH + H+ | - |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | glycolate + NAD+ | - |
r | |
glyoxylate + NADPH + H+ | - |
Thermus thermophilus HB27 | glycolate + NADP+ | - |
ir | |
additional information | phenylpyruvate, pyruvate, methylglyoxal, malate, and oxaloacetate not suitable as substrate | Thermus thermophilus HB27 | ? | - |
? | |
additional information | substrate specificity, several 2-oxo compounds including phenylpyruvate, pyruvate, methylglyoxal, and oxaloacetate act as inert electron acceptors, as do glycolate and malate, overview | Thermus thermophilus | ? | - |
? | |
additional information | substrate specificity, several 2-oxo compounds including phenylpyruvate, pyruvate, methylglyoxal, and oxaloacetate act as inert electron acceptors, as do glycolate and malate, overview | Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 36000, SDS-PAGE, native mass by gel filtration (native mass not given in the reference) | Thermus thermophilus HB27 |
Synonyms | Comment | Organism |
---|---|---|
More | cf. 1.1.1.26 | Thermus thermophilus |
TthGR1 | belongs to the D-2-hydroxyacid dehydrogenase family, enzyme used NADPH and NADH (EC 1.1.1.26 activity) as cofactor | Thermus thermophilus HB27 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
recombinant enzyme | Thermus thermophilus |
45 | - |
recombinant protein | Thermus thermophilus HB27 |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
10 min, recombinant enzyme, stable | Thermus thermophilus |
70 | - |
stable up to 70°C | Thermus thermophilus HB27 |
80 | - |
complete loss of activity within 10 min when incubates above 80°C | Thermus thermophilus HB27 |
82 | - |
melting point | Thermus thermophilus HB27 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
22000 | - |
NADH | pH 6.7, 45°C, substrate glyoxylate, purified recombinant enzyme | Thermus thermophilus | |
30000 | - |
NADPH | pH 6.7, 45°C, substrate glyoxylate, purified recombinant enzyme | Thermus thermophilus | |
65000 | - |
glyoxylate | pH 6.7, 45°C, cofactor NADPH, purified recombinant enzyme | Thermus thermophilus | |
76000 | - |
glyoxylate | pH 6.7, 45°C, cofactor NADH, purified recombinant enzyme | Thermus thermophilus | |
220000 | - |
NADH | with glyoxylate as substrate, pH 6.7, 45°C | Thermus thermophilus HB27 | |
300000 | - |
NADPH | with glyoxylate as substrate, pH 6.7, 45°C | Thermus thermophilus HB27 | |
650000 | - |
glyoxylate | with as NADPH as cofactor, pH 6.7, 45°C | Thermus thermophilus HB27 | |
760000 | - |
glyoxylate | with as NADH as cofactor, pH 6.7, 45°C | Thermus thermophilus HB27 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.7 | - |
recombinant enzyme | Thermus thermophilus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.7 | - |
recombinant protein | Thermus thermophilus HB27 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | less active than NADPH | Thermus thermophilus | |
NADH | yields higher Km but similar turnover than NADPH | Thermus thermophilus HB27 | |
NADPH | is the preferred cofactor | Thermus thermophilus | |
NADPH | preferred cofactor, yields lower Km but similar turnover than NADH | Thermus thermophilus HB27 |