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Literature summary for 1.1.1.79 extracted from

  • Ogino, H.; Nakayama, H.; China, H.; Kawata, T.; Doukyu, N.; Yasuda, M.
    Characterization of recombinant glyoxylate reductase from thermophile Thermus thermophilus HB27 (2008), Biotechnol. Prog., 24, 321-325.
    View publication on PubMed

Cloned(Commentary)

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

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 [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 [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/ Products (Substrates)

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 Stability

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

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-
Thermus thermophilus HB27
-
NBRC code 101085
-
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
-
-

Purification (Commentary)

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 and Products (Substrate)

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

Subunits Comment Organism
dimer 2 * 36000, SDS-PAGE, native mass by gel filtration (native mass not given in the reference) Thermus thermophilus HB27

Synonyms

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
recombinant enzyme Thermus thermophilus
45
-
recombinant protein Thermus thermophilus HB27

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.7
-
recombinant enzyme Thermus thermophilus

pH Range

pH Minimum pH Maximum Comment Organism
6.7
-
recombinant protein Thermus thermophilus HB27

Cofactor

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