Cloned (Comment) | Organism |
---|---|
gene guaB, recombinant expresion of HIs8-tagged wild-type and mutant enzymes in Escherichia coli | Eremothecium gossypii |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant apoenzyme AgIMPDH-DELTABateman, vapor diffusion method, mixing 23 mg/ml protein 10 mM Tris-HCl, 100 mM KCl, 0.25 mM TCEP, pH 8.0, with an equal volume of mother liquor consisting of 0.1 M HEPES, pH 7.5, 40% PEG 300 v/v, and 0.2 M NaCl, for the enzyme mutant complex with IMP and NAD+ up to 5 mM XMP and 5 mM NAD+ are added to the protein solution, at room temperature, harvest of crystals of AgIMPDH-DELTABateman with a mixture of about 80% of the covalent intermediate E-XMP* and about 20% IMP bound to the active site, X-ray diffraction structure determination and analysis | Eremothecium gossypii |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of the catalytically active AgIMPDH-DELTABateman enzyme mutant, that has a the deletion of the regulatory domain facilitating crystallization | Eremothecium gossypii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Mycophenolic acid | - |
Eremothecium gossypii | |
NAD+ | substrate inhibition | Eremothecium gossypii | |
XMP | product inhibition | Eremothecium gossypii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten enzyme kinetics | Eremothecium gossypii | |
0.0902 | - |
IMP | recombinant wild-type enzyme, pH 8.0, 28°C | Eremothecium gossypii | |
0.118 | - |
IMP | recombinant AgIMPDH-DELTABateman mutant enzyme, pH 8.0, 28°C | Eremothecium gossypii | |
0.28 | - |
NAD+ | recombinant wild-type enzyme, pH 8.0, 28°C | Eremothecium gossypii | |
0.387 | - |
NAD+ | recombinant AgIMPDH-DELTABateman mutant enzyme, pH 8.0, 28°C | Eremothecium gossypii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
IMP + NAD+ + H2O | Eremothecium gossypii | - |
XMP + NADH + H+ | - |
? | |
IMP + NAD+ + H2O | Eremothecium gossypii ATCC 10895 | - |
XMP + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Eremothecium gossypii | Q756Z6 | single gene | - |
Eremothecium gossypii ATCC 10895 | Q756Z6 | single gene | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
IMP + NAD+ + H2O = XMP + NADH + H+ | reaction in two steps via covalent interemediate state, the catalytic residue is Cys334 | Eremothecium gossypii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
IMP + NAD+ + H2O | - |
Eremothecium gossypii | XMP + NADH + H+ | - |
? | |
IMP + NAD+ + H2O | - |
Eremothecium gossypii ATCC 10895 | XMP + NADH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the core of the catalytic domain comprises a (beta/alpha)8 barrel which represents the typical triose-phosphate isomerase fold (TIM barrel) | Eremothecium gossypii |
tetramer | enzyme structure in solution | Eremothecium gossypii |
Synonyms | Comment | Organism |
---|---|---|
IMP dehydrogenase | - |
Eremothecium gossypii |
IMPDH | - |
Eremothecium gossypii |
inosine 5-monophosphate dehydrogenase | - |
Eremothecium gossypii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | - |
assay at | Eremothecium gossypii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.46 | - |
IMP | recombinant wild-type enzyme, pH 8.0, 28°C | Eremothecium gossypii | |
3.06 | - |
IMP | recombinant AgIMPDH-DELTABateman mutant enzyme, pH 8.0, 28°C | Eremothecium gossypii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Eremothecium gossypii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme shows a clear preference for NAD+ over NADP+, since it loses more than 95% activity when using the latter as co-substrate | Eremothecium gossypii | |
NAD+ | - |
Eremothecium gossypii |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000321 | - |
Mycophenolic acid | pH 8.0, 28°C, recombinant enzyme | Eremothecium gossypii | |
0.2083 | - |
XMP | pH 8.0, 28°C, recombinant enzyme | Eremothecium gossypii | |
3.6 | - |
NAD+ | recombinant wild-type enzyme, pH 8.0, 28°C | Eremothecium gossypii | |
4.5 | - |
NAD+ | recombinant AgIMPDH-DELTABateman mutant enzyme, pH 8.0, 28°C | Eremothecium gossypii |
General Information | Comment | Organism |
---|---|---|
malfunction | overexpression of the IMPDH gene increases the metabolic flux through the guanine pathway and ultimately enhances 40% riboflavin production with respect to the wild-type. IMPDH disruption results in a 100fold increase of inosine excretion to the culture media | Eremothecium gossypii |
metabolism | the enzyme inosine-5'-monophosphate dehydrogenase (IMPDH) catalyzes the rate-limiting step in the guanine nucleotide de novo biosynthetic pathway and plays a key role in controlling the cellular nucleotide pools. The enzyme, UniProt ID AER117W, is the only one responsible for the IMPDH activity in Ashbya gossypiis | Eremothecium gossypii |
additional information | Ashbya gossypii is a natural riboflavin superproducer | Eremothecium gossypii |
physiological function | the enzyme inosine-5'-monophosphate dehydrogenase (IMPDH) catalyzes the rate-limiting step in the guanine nucleotide de novo biosynthetic pathway and plays a key role in controlling the cellular nucleotide pools. IMPDH is an important metabolic bottleneck in the guanine nucleotide synthesis. Guanine nucleotides are essential for cell viability | Eremothecium gossypii |