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Literature summary for 1.2.1.9 extracted from

  • Ettema, T.J.; Ahmed, H.; Geerling, A.C.; van der Oost, J.; Siebers, B.
    The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) of Sulfolobus solfataricus: a key-enzyme of the semi-phosphorylative branch of the Entner-Doudoroff pathway. (2007), Extremophiles, 12, 75-88.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
6-O-phosphono-D-fructofuranose minor stimulatory effect Saccharolobus solfataricus
AMP minor stimulatory effect Saccharolobus solfataricus
D-fructose minor stimulatory effect Saccharolobus solfataricus
D-glucose 1-phosphate 14fold activation at 0.1 mM D-glucose 1-phosphate Saccharolobus solfataricus
pyruvate minor stimulatory effect Saccharolobus solfataricus

Cloned(Commentary)

Cloned (Comment) Organism
cloned and expressed in BL21 (DE3) CodonPlus using the pET expression vector system Saccharolobus solfataricus
expressed in Escherichia coli BL21(DE3) cells Saccharolobus solfataricus
expressed in Escherichia coli BL21(DE3) CodonPlus cells Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
ATP slight inhibition Saccharolobus solfataricus
galactonate slight inhibition; slight inhibition (1.2 to 1.3fold); slight inhibition (1.2 to 1.3fold) Saccharolobus solfataricus
gluconate slight inhibition Saccharolobus solfataricus
additional information glyceraldehyde and ADP have no effect on GAPN activity; not inhibited by glyceraldehyde and ADP; not inhibited by glyceraldehyde and ADP Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.09
-
NADP+ 70┬░C, pH 7 Saccharolobus solfataricus
0.09
-
NADP+ in 90 mM HEPES/KOH, 160 mM KCl, pH 7.0, at 70┬░C Saccharolobus solfataricus
0.09
-
NADP+ in the absence or presence of D-glucose 1-phosphate Saccharolobus solfataricus
0.501
-
DL-glyceraldehyde 3-phosphate in the presence of D-glucose 1-phosphate Saccharolobus solfataricus
0.51
-
DL-glyceraldehyde 3-phosphate 70┬░C, pH 7 Saccharolobus solfataricus
0.51
-
D-Glyceraldehyde-3-phosphate in 90 mM HEPES/KOH, 160 mM KCl, pH 7.0, at 70┬░C Saccharolobus solfataricus
0.51
-
DL-glyceraldehyde 3-phosphate in the absence of D-glucose 1-phosphate Saccharolobus solfataricus
17.4
-
NAD+ 70┬░C, pH 7 Saccharolobus solfataricus
17.4
-
NAD+ in 90 mM HEPES/KOH, 160 mM KCl, pH 7.0, at 70┬░C Saccharolobus solfataricus
17.4
-
NAD+ in the absence of D-glucose 1-phosphate Saccharolobus solfataricus
21.1
-
NAD+ in the presence of D-glucose 1-phosphate Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56930
-
subunit, calculated from amino acid sequence Saccharolobus solfataricus
57000
-
subunit, SDS-PAGE Saccharolobus solfataricus
57000
-
x * 57000, SDS-PAGE Saccharolobus solfataricus
57000
-
4 * 57000, SDS-PAGE Saccharolobus solfataricus
189000
-
-
Saccharolobus solfataricus
189000
-
native enzyme Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-glyceraldehyde 3-phosphate + NADP+ + H2O Saccharolobus solfataricus the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase is a key-enzyme of the semi-phosphorylative branch of the Entner┬ľDoudoroff pathway 3-phospho-D-glycerate + NADPH + 2 H+
-
ir

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-
Saccharolobus solfataricus Q97U30
-
-
Saccharolobus solfataricus Q97XA5 isozyme Gap-N2
-
Saccharolobus solfataricus Q97XS9 isozyme GapN-1
-
Saccharolobus solfataricus P2 Q97U30
-
-
Saccharolobus solfataricus P2 Q97XA5 isozyme Gap-N2
-
Saccharolobus solfataricus P2 Q97XS9 isozyme GapN-1
-

Purification (Commentary)

Purification (Comment) Organism
heat precipitation and Superdex 200 gel filtration Saccharolobus solfataricus
Superdex 200 gel filtration Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
additional information
-
the recombinant enzyme has a specific activity of 3.74 U/mg Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glyceraldehyde 3-phosphate + NAD+ + H2O enzyme shows negligible activity with NAD+. The NAD+-dependent reaction shows no saturation at NAD+ concentrations up to 50 mM. The highest enzyme activity is observed at 50 mM NAD+ which is 7fold to 8fold lower than the Vmax observed using NADP+ as cofactor Saccharolobus solfataricus 3-phospho-D-glycerate + NADH + H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Saccharolobus solfataricus 3-phospho-D-glycerate + NADPH + H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase is a key-enzyme of the semi-phosphorylative branch of the Entner┬ľDoudoroff pathway Saccharolobus solfataricus 3-phospho-D-glycerate + NADPH + 2 H+
-
ir
D-glyceraldehyde-3-phosphate + NAD+ weak reaction Saccharolobus solfataricus D-3-phosphoglycerate + NADH + H+
-
ir
D-glyceraldehyde-3-phosphate + NAD+ weak reaction Saccharolobus solfataricus P2 D-3-phosphoglycerate + NADH + H+
-
ir
D-glyceraldehyde-3-phosphate + NADP+
-
Saccharolobus solfataricus D-3-phosphoglycerate + NADPH + H+
-
ir
D-glyceraldehyde-3-phosphate + NADP+
-
Saccharolobus solfataricus P2 D-3-phosphoglycerate + NADPH + H+
-
ir
DL-glyceraldehyde 3-phosphate + NAD+ + H2O NADP+ is much more efficient than NAD+ Saccharolobus solfataricus 3-phospho-DL-glycerate + NADH + H+
-
ir
DL-glyceraldehyde 3-phosphate + NAD+ + H2O NADP+ is much more efficient than NAD+ Saccharolobus solfataricus P2 3-phospho-DL-glycerate + NADH + H+
-
ir
DL-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Saccharolobus solfataricus 3-phospho-DL-glycerate + NADPH + H+
-
ir
DL-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Saccharolobus solfataricus P2 3-phospho-DL-glycerate + NADPH + H+
-
ir
DL-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Saccharolobus solfataricus 3-phospho-D-glycerate + NADPH + H+
-
r
additional information the enzyme shows only negligible activity with NAD+ Saccharolobus solfataricus ?
-
?
additional information the enzyme shows only negligible activity with NAD+ Saccharolobus solfataricus P2 ?
-
?

Subunits

Subunits Comment Organism
? x * 57000, SDS-PAGE Saccharolobus solfataricus
homotetramer 4 * 57000, SDS-PAGE Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
GAP dehydrogenase
-
Saccharolobus solfataricus
GAPN
-
Saccharolobus solfataricus
non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase
-
Saccharolobus solfataricus
non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase GAPN
-
Saccharolobus solfataricus

Cofactor

Cofactor Comment Organism Structure
NAD+ enzyme shows negligible activity with NAD+. The NAD+-dependent reaction of Sso-GAPN shows no saturation at NAD+ concentrations up to 50 mM. The highest enzyme activity is observed at 50 mM NAD+ which is 7fold to 8fold lower than the Vmax observed using NADP+ as cofactor Saccharolobus solfataricus
NAD+ negligible activity with NAD+ Saccharolobus solfataricus
NADP+
-
Saccharolobus solfataricus
NADP+ prefers NADP+ more than NAD+ as cofactor Saccharolobus solfataricus
NADP+ the enzyme shows only negligible activity with NAD+ compared to the physiological cosubstrate NADP+ Saccharolobus solfataricus