Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Rasamsonia emersonii |
expression of histidine-tagged wild type Texr and mutant Texr K271R/N273D in Escherichia coli BL21-Star DE3 | Rasamsonia emersonii |
Protein Variants | Comment | Organism |
---|---|---|
K271R/N273D | in the double mutant, affinity for NADPH decreases 3.1fold, while affinity for NADH remains relatively unchanged in comparison with the wild-type enzyme. The turnover number increases 1.6fold for the double mutant with NADH and decreases 3.2fold with NADPH relative to the wild-type enzyme. As a consequence, the catalytic efficiency of the double mutant (kcat/Km) increases 1.4fold with NADH and decreases 10.8fold with NADPH relative to the wild-type enzyme. Using the specificity constant (kcat/Km (NADH)/kcat/Km(NADPH)) the coenzyme preference for NADH is improved 16fold in the TeXR K271R/N273D double-mutant enzyme | Rasamsonia emersonii |
K271R/N273D | catalytic efficiency of the double mutant increases 1.4fold with NADH and decreases 10.8fold with NADPH relative to the wild-type enzyme | Rasamsonia emersonii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.244 | - |
NADPH | pH 6.5, 37°C, wild-type enzyme | Rasamsonia emersonii | |
0.244 | - |
NADPH | wild-type, pH 6.5, 37°C | Rasamsonia emersonii | |
0.263 | - |
NADH | pH 6.5, 37°C, wild-type enzyme | Rasamsonia emersonii | |
0.263 | - |
NADH | wild-type, pH 6.5, 37°C | Rasamsonia emersonii | |
0.3 | - |
NADH | pH 6.5, 37°C, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
0.3 | - |
NADH | mutant K271R/N273, pH 6.5, 37°C | Rasamsonia emersonii | |
0.747 | - |
NADPH | pH 6.5, 37°C, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
0.747 | - |
NADPH | mutant K271R/N273, pH 6.5, 37°C | Rasamsonia emersonii | |
24.56 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADPH, wild-type enzyme | Rasamsonia emersonii | |
24.6 | - |
D-xylose | wild-type, cosubstrate NADPH, pH 6.5, 37°C | Rasamsonia emersonii | |
76.06 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADH, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
76.1 | - |
D-xylose | mutant K271R/N273, cosubstrate NADH, pH 6.5, 37°C | Rasamsonia emersonii | |
76.5 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADPH, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
76.5 | - |
D-xylose | mutant K271R/N273, cosubstrate NADPH, pH 6.5, 37°C | Rasamsonia emersonii | |
89.4 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADH, wild-type enzyme | Rasamsonia emersonii | |
89.4 | - |
D-xylose | wild-type, cosubstrate NADH, pH 6.5, 37°C | Rasamsonia emersonii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36000 | - |
x * 36000, calculated, x * 40000, SDS-PAGE | Rasamsonia emersonii |
40000 | - |
x * 40000, wild-type and double-mutant (K271R/N273D) histidine-tagged protein, SDS-PAGE | Rasamsonia emersonii |
40000 | - |
x * 36000, calculated, x * 40000, SDS-PAGE | Rasamsonia emersonii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylose + NADPH + H+ | Rasamsonia emersonii | expression of Texr is inducible by the same carbon sources responsible for the induction of genes encoding enzymes relevant to lignocellulose hydrolysis, suggesting a coordinated expression of intracellular and extracellular enzymes relevant to hydrolysis and metabolism of pentose sugars in Talaromyces emersonii in adaptation to its natural habitat. This indicates a potential advantage in survival and response to a nutrient-poor environment | xylitol + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rasamsonia emersonii | C5J3R6 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Rasamsonia emersonii |
recombinant protein | Rasamsonia emersonii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-xylose + NADH + H+ | - |
Rasamsonia emersonii | xylitol + NAD+ | - |
? | |
D-xylose + NADH + H+ | wild-type TeXR shows dual coenzyme specificity but is preferentially NADPH-dependent, with affinity for NADPH being 1.1fold higher than NADH and catalytic efficiency (kcat/Km) 24.5fold higher with NADPH as coenzyme. Affinity for xylose is 3.6fold higher with NADPH as coenzyme | Rasamsonia emersonii | xylitol + NAD+ | - |
? | |
D-xylose + NADPH + H+ | - |
Rasamsonia emersonii | xylitol + NADP+ | - |
? | |
D-xylose + NADPH + H+ | expression of Texr is inducible by the same carbon sources responsible for the induction of genes encoding enzymes relevant to lignocellulose hydrolysis, suggesting a coordinated expression of intracellular and extracellular enzymes relevant to hydrolysis and metabolism of pentose sugars in Talaromyces emersonii in adaptation to its natural habitat. This indicates a potential advantage in survival and response to a nutrient-poor environment | Rasamsonia emersonii | xylitol + NADP+ | - |
? | |
D-xylose + NADPH + H+ | wild-type TeXR shows dual coenzyme specificity but is preferentially NADPH-dependent, with affinity for NADPH being 1.1fold higher than NADH and catalytic efficiency (kcat/Km) 24.5fold higher with NADPH as coenzyme. Affinity for xylose is 3.6fold higher with NADPH as coenzyme | Rasamsonia emersonii | xylitol + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 40000, wild-type and double-mutant (K271R/N273D) histidine-tagged protein, SDS-PAGE | Rasamsonia emersonii |
? | x * 36000, calculated, x * 40000, SDS-PAGE | Rasamsonia emersonii |
Synonyms | Comment | Organism |
---|---|---|
Texr | - |
Rasamsonia emersonii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Rasamsonia emersonii |
37 | - |
wild-type and double-mutant (K271R/N273D) protein | Rasamsonia emersonii |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 60 | active from 30°C to 60°C, wild-type and double-mutant (K271R/N273D) protein | Rasamsonia emersonii |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
retained 28% of activity at 60°C | Rasamsonia emersonii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15750 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADH, wild-type enzyme | Rasamsonia emersonii | |
15750 | - |
NADH | wild-type, pH 6.5, 37°C | Rasamsonia emersonii | |
25110 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADH, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
25110 | - |
NADH | mutant K271R/N273, pH 6.5, 37°C | Rasamsonia emersonii | |
100900 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADPH, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
100900 | - |
NADPH | mutant K271R/N273, pH 6.5, 37°C | Rasamsonia emersonii | |
324000 | - |
D-xylose | pH 6.5, 37°C, cosubstrate: NADPH, wild-type enzyme | Rasamsonia emersonii | |
324000 | - |
NADPH | wild-type, pH 6.5, 37°C | Rasamsonia emersonii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
- |
Rasamsonia emersonii |
6.5 | - |
wild-type and double-mutant (K271R/N273D) protein | Rasamsonia emersonii |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | - |
Rasamsonia emersonii |
5 | 8 | active from pH 5 to pH 8, wild-type and double-mutant (K271R/N273D) protein | Rasamsonia emersonii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | catalytic efficiency is 24.5fold higher with NADPH as coenzyme than with NADH | Rasamsonia emersonii | |
NADPH | wild-type TeXR shows dual coenzyme specificity but is preferentially NADPH-dependent, with affinity for NADPH being 1.1fold higher than NADH and catalytic efficiency (kcat/Km) 24.5fold higher with NADPH as coenzyme. Affinity for xylose is 3.6fold higher with NADPH as coenzyme. K271R/N273D double mutant displays an altered coenzyme preference with a 16fold improvement in NADH utilization relative to the wild type | Rasamsonia emersonii | |
NADPH | catalytic efficiency is 24.5fold higher with NADPH as coenzyme than with NADH. Affinity for xylose is 3.6fold higher with NADPH as coenzyme | Rasamsonia emersonii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
59920 | - |
NADH | pH 6.5, 37°C, wild-type enzyme | Rasamsonia emersonii | |
59920 | - |
NADH | wild-type, pH 6.5, 37°C | Rasamsonia emersonii | |
83750 | - |
NADH | pH 6.5, 37°C, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
83750 | - |
NADH | mutant K271R/N273, pH 6.5, 37°C | Rasamsonia emersonii | |
135500 | - |
NADPH | pH 6.5, 37°C, mutant enzyme K271R/N273D | Rasamsonia emersonii | |
135500 | - |
NADPH | mutant K271R/N273, pH 6.5, 37°C | Rasamsonia emersonii | |
1466000 | - |
NADPH | pH 6.5, 37°C, wild-type enzyme | Rasamsonia emersonii | |
1466000 | - |
NADPH | wild-type, pH 6.5, 37°C | Rasamsonia emersonii |