Cloned (Comment) | Organism |
---|---|
production of the beta subunit in Escherichia coli | Azospirillum brasilense |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | for the NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme, uncompetitive inhibition with ferricyanide or iodonitrotetrazolium as substrate, competitive inhibition with NADPH as substrate | Azospirillum brasilense |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0035 | - |
NADPH | of NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme, acceptor: iodonitrotetrazolium | Azospirillum brasilense | |
0.0098 | - |
NADPH | of NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme, acceptor: menadione | Azospirillum brasilense | |
0.0118 | - |
NADPH | of NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme, acceptor: ferricyanide | Azospirillum brasilense | |
0.035 | - |
menadione | of NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme | Azospirillum brasilense | |
0.05 | - |
iodonitrotetrazolium | of NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme | Azospirillum brasilense | |
0.14 | - |
ferricyanide | of NADPH: acceptor oxidoreductase activity of the beta subunit of the enzyme | Azospirillum brasilense |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | iron-sulfur protein | Azospirillum brasilense |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Azospirillum brasilense | - |
- |
- |
Purification (Comment) | Organism |
---|---|
of the recombinant enzyme beta subunit, using ammonium sulfate fractionation, affinity chromatography on Reactive Red, ultrafiltration and column chromatography on Ultrogel AcA 54 | Azospirillum brasilense |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ferricyanide + NADPH + H+ | - |
Azospirillum brasilense | ferrocyanide + NADP+ | - |
r | |
iodonitrotetrazolium + NADPH + H+ | - |
Azospirillum brasilense | reduced iodonitrotetrazolium + NADP+ | - |
r | |
L-glutamine + 2-oxoglutarate + NADPH + H+ | - |
Azospirillum brasilense | L-glutamate + NADP+ | - |
? | |
menadione + NADPH + H+ | - |
Azospirillum brasilense | menadiol + NADP+ | - |
r | |
additional information | the enzyme beta subunit is devoid of glutamate synthase activity in either direction at both pH 7.5 and 9.5, but it can oxidize NADPH and transfer electrons to synthetic electron acceptors like iodonitrotetrazolium, ferricyanide, menadione, dichloroindophenol, the beta subunit is highly specific toward NADPH, the rate of oxidation of NADH in the presence of electron acceptors is less than 5% of that measured with NADPH | Azospirillum brasilense | ? | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
18.3 | - |
NADPH | of NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, co-substrate: iodonitrotetrazolium, inhibitor: 2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | Azospirillum brasilense | |
20 | - |
iodonitrotetrazolium | of NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, co-substrate: NADPH, inhibitor: 2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | Azospirillum brasilense | |
25.3 | - |
menadione | of NADPH:acceptor oxidoreductase acitivity of the beta subunit of the enzyme | Azospirillum brasilense | |
30.8 | - |
NADPH | of NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, co-substrate: ferricyanide, inhibitor: 2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | Azospirillum brasilense | |
32 | - |
iodonitrotetrazolium | of NADPH:acceptor oxidoreductase acitivity of the beta subunit of the enzyme | Azospirillum brasilense | |
39.5 | - |
ferricyanide | of NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, co-substrate: NADPH, inhibitor: 2'-phosphoadenosine-5'-diphospho-5'beta-D-ribose | Azospirillum brasilense | |
41.9 | - |
ferricyanide | of NADPH:acceptor oxidoreductase acitivity of the beta subunit of the enzyme | Azospirillum brasilense |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
flavin | flavoenzyme | Azospirillum brasilense | |
flavin | the beta subunit contains the binding site for FAD, 0.83 mol FAD per mol beta subunit | Azospirillum brasilense | |
additional information | the enzyme contains three distinct ion-sulfur centers per alphabeta protomer | Azospirillum brasilense | |
NADPH | the beta subunit contains the NADPH binding site of the enzyme | Azospirillum brasilense |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.021 | - |
2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | for the NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, substrate: NADPH, cosubstrate: ferricyanide | Azospirillum brasilense | |
0.08 | - |
2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | for the NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, substrate: NADPH, cosubstrate: iodonitrotetrazolium | Azospirillum brasilense | |
0.18 | - |
2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | for the NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, substrate: ferricyanide, cosubstrate: NADPH | Azospirillum brasilense | |
0.495 | - |
2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose | for the NADPH:acceptor oxidoreductase activity of the beta subunit of the enzyme, substrate: iodonitrotetrazolium, cosubstrate: NADPH | Azospirillum brasilense |