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Literature summary for 1.5.1.40 extracted from

  • Le, C.; Oyugi, M.; Joseph, E.; Nguyen, T.; Ullah, M.; Aubert, J.; Phan, T.; Tran, J.; Johnson-Winters, K.
    Effects of isoleucine 135 side chain length on the cofactor donor-acceptor distance within F420H2 NADP+ oxidoreductase A kinetic analysis (2017), Biochem. Biophys. Rep., 9, 114-120 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.1.40

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C41(DE3) Archaeoglobus fulgidus

Protein Variants

Protein Variants Comment Organism
I135A site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Archaeoglobus fulgidus
I135G site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Archaeoglobus fulgidus
I135V site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Archaeoglobus fulgidus
additional information pre-steady-state data with F420 cofactor and NADPH for the enzyme Fno mutant variants reveal biphasic kinetics with a fast and slow phase, similar with wild-type Fno, overview Archaeoglobus fulgidus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information substrate binding studies, steady-state and pre steady-state kinetic analysis with wild-type enzyme Fno and Ile135 Fno mutant variants, I135A, I135V, and I135G, overview. Steady-state kinetic analysis of wild-type Fno and the variants show classical Michaelis-Menten kinetics with varying FO concentrations. The data reveal a decreased kcat as side chain length decreased, with varying FO concentrations. The steady-state plots reveal non-Michaelis-Menten kinetic behavior when NADPH is varied. The double reciprocal plot of the varying NADPH concentrations displays a downward concave shape, while the NADPH binding curves gave Hill coefficients of less than 1. These data suggest that negative cooperativity occurs between the two identical monomers. The pre steady-state Abs420 versus time trace reveals biphasic kinetics, with a fast phase (hydride transfer) and a slow phase. The fast phase displays an increased rate constant as side chain length decreases. The rate constant for the second phase, remained about 2/s for each variant. Pre-steady-state data with F420 cofactor and NADPH for the enzyme Fno mutant variants reveal biphasic kinetics with a fast and slow phase, similar with wild-type Fno, overview Archaeoglobus fulgidus
0.00027
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
0.0007
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
0.0023
-
 NADPH phase I, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
0.0029
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
0.0036
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
0.0036
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
0.0037
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
0.004
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
0.016
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
0.051
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
0.062
-
 NADPH phase II, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
0.654
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
reduced coenzyme F420 + NADP+ Archaeoglobus fulgidus
-
 oxidized coenzyme F420 + NADPH + H+
-
 r

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus O29370
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from cell-free extracts of Escherichia coli strain C41(DE3) by heat treatment at 90°C for 30 min, ammonium sulfate fractionation with addition of 0.05% polyethylenimine, followed by an ion exchange chromatography, ultrafiltration, and gel filtration Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information effects of side chain length of residue Il135 on the donor-acceptor distance between NADP+ and the F420 precursor, FO, overview Archaeoglobus fulgidus ?
-
 ?
reduced coenzyme F420 + NADP+
-
 Archaeoglobus fulgidus oxidized coenzyme F420 + NADPH + H+
-
 r

Synonyms

Synonyms Comment Organism
F420H2:NADP+ oxidoreductase
-
 Archaeoglobus fulgidus
Fno
-
 Archaeoglobus fulgidus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at Archaeoglobus fulgidus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.11
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
0.33
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
0.7
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
0.91
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
1.24
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
1.5
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
1.6
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
1.8
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
2.16
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
4.16
-
 NADPH phase I, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
5.3
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
5.41
-
 NADPH phase II, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 assay at Archaeoglobus fulgidus

General Information

General Information Comment Organism
physiological function residue I135 plays a key role in sustaining the donor-acceptor distance between the two cofactor substrates, thereby regulating the rate at which the hydride is transferred from FOH2 to NADP+. Fno is a dynamic enzyme that regulates NADPH production Archaeoglobus fulgidus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.5
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
6.8
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
42
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
88
-
 NADPH phase II, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
194.4
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135G Archaeoglobus fulgidus
420
-
 NADPH phase II, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
444.4
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus
486.5
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
1325
-
 oxidized coenzyme F420 with F420 precursor, FO, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
1800
-
 NADPH phase I, pH 6.5, 22°C, recombinant wild-type enzyme Archaeoglobus fulgidus
2100
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135V Archaeoglobus fulgidus
3400
-
 NADPH phase I, pH 6.5, 22°C, recombinant mutant I135A Archaeoglobus fulgidus