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Information on EC 1.2.1.89 - D-glyceraldehyde dehydrogenase (NADP+) and Organism(s) Thermoplasma acidophilum and UniProt Accession Q9HK01

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IUBMB Comments
The enzyme from the archaea Thermoplasma acidophilum and Picrophilus torridus is involved in the non-phosphorylative Entner-Doudoroff pathway. cf. EC 1.2.99.8, glyceraldehyde dehydrogenase (FAD-containing).
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Thermoplasma acidophilum
UNIPROT: Q9HK01
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The taxonomic range for the selected organisms is: Thermoplasma acidophilum
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
glyceraldehyde dehydrogenase, ta0809, pto0332, more
SYSTEMATIC NAME
IUBMB Comments
D-glyceraldehyde:NADP+ oxidoreductase
The enzyme from the archaea Thermoplasma acidophilum and Picrophilus torridus is involved in the non-phosphorylative Entner-Doudoroff pathway. cf. EC 1.2.99.8, glyceraldehyde dehydrogenase (FAD-containing).
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-glyceraldehyde + NAD+
D-glycerate + NADH + H+
show the reaction diagram
NAD+ is a very poor cofactor for wild-type
-
-
?
D-glyceraldehyde + NADP+ + H2O
D-glycerate + NADPH + H+
show the reaction diagram
D-glyceraldehyde 3-phosphate + NADP+ + H2O
D-glycerate 3-phosphate + NADPH + H+
show the reaction diagram
glyceraldehyde + NAD+ + H2O
glycerate + NADH + H+
show the reaction diagram
-
-
-
?
glycolaldehyde + NADP+
glycolate + NADPH + H+
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
D-glyceraldehyde + NADP+ + H2O
D-glycerate + NADPH + H+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
poor cofoactor
NADP+
NADPH
NADPH is preferred over NADH
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
1 mM, 12 h, 4°C, complete loss of activity
Cd2+
1 mM, 12 h, 4°C, 93% loss of activity
Cu2+
1 mM, 12 h, 4°C, complete loss of activity
Hg2+
1 mM, 12 h, 4°C, 99% loss of activity
Mg2+
1 mM, 12 h, 4°C, 24% loss of activity
Mn2+
1 mM, 12 h, 4°C, 44% loss of activity
Ni2+
1 mM, 12 h, 4°C, 58% loss of activity
Zn2+
1 mM, 12 h, 4°C, 46% loss of activity
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
1 mM, activity is enhanced more than 2fold
4-chloromercuribenzoic acid
1 mM, completely inhibits activity
4-hydroxymercuribenzoic acid
1 mM, completely inhibits activity
DTT
1 mM, activity is enhanced more than 2fold
glutathione
1 mM, activity is enhanced more than 2fold
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2 - 0.33
D-glyceraldehyde
18.3
D-glycerate
pH 6.9, 58°C
12.3
glycolaldehyde
pH 6.9, 58°C
16.7 - 22.7
NAD+
0.017 - 0.36
NADP+
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
26
D-glyceraldehyde
pH 6.9, 58°C
20
D-glycerate
pH 6.9, 58°C
37
glycolaldehyde
pH 6.9, 58°C
1.64 - 10.99
NAD+
25
NADP+
pH 6.9, 58°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
130
D-glyceraldehyde
pH 6.9, 58°C
1.1
D-glycerate
pH 6.9, 58°C
3
glycolaldehyde
pH 6.9, 58°C
0.07 - 0.63
NAD+
70
NADP+
pH 6.9, 58°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 8.7
pH 5.8: about 50% of maximal activity, pH 8.7: about 50% of maximal activity
6 - 10
pH 6.0: about 65% of maximal activity, pH 10.0: about 70% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 70
30°C: about 50% of maximal activity, 70°C: about 60% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
120000
gel filtration
215000
gel filtration
53000
4 * 53000, SDS-PAGE
54700
2 * 54700, calculated from sequence
54782
4 * 54782, calculated from sequence
55000
x * 55000, SDS-PAGE
56000
2 * 56000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homotetramer
4 * 53000, SDS-PAGE
tetramer
4 * 54782, calculated from sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structures of wild-type and mutant F34M/S405N, to 1.95 A resolution for wild-type and 2.14 and 2.10 A resolution for the mutant
wild type enzyme, sitting drop vapor diffusion method, using 50 mM MOPS, 50 mM sodium HEPES pH 7.5, 20% (w/v) ethylene glycol, 10% (w/v) PEG 8000, 20 mM glutamic acid (racemic), 20 mM glycine, 20 mM serine (racemic), 20 mM alanine (racemic), 20 mM lysine-HCl (racemic). Mutant enzyme F34M/S405N, sitting drop vapor diffusion method, using 50 mM MES, 50 mM imidazole pH 6.5, 8.3% (w/v) PEG 8000, 16.7% (w/v) ethylene glycol, 20 mM sodium formate, 20 mM sodium citrate, 20 mM sodium oxamate, 20 mM ammonium acetate, 20 mM sodium potassium tartrate
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34M/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 55.7
F34L
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 3.2
F34M/D372N/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 7.2
F34M/D372N/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 35.1
F34M/S405N
F34M/W271R/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 22.6
F34M/W271S/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 42.5
F34M/W271S/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 1.1
F34M/Y399C/S405N
enhances enzyme activity with the cofactor NAD+ by a factor of eight and enhances solubilty
S405C
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 6
S405N
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 7.4
W271S
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 17.1
Y399C
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
24 h, stable
724216
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70
half-life: 40 min
80
activity is decreased to less than 50% within 20 min
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
guanidine-HCl
the stability of the enzyme is ca. 28 kJ/mol both at 25°C and 40°C
isobutanol
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography
HiTrap column chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
the gene of TaAlDH is synthesized and cloned into a T7 based vector system for recombinant expression in Escherichia coli. Using matrix-assisted refolding of inclusion bodies the yield of enzyme production is enhanced 43fold
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
the enzyme accept NAD+ as cofactor under technically relevant conditions, making it suitable for application in the artificial glycolysis by synthetic cascade biomanufacturing
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Jung, J.H.; Lee, S.B.
Identification and characterization of Thermoplasma acidophilum glyceraldehyde dehydrogenase: a new class of NADP+-specific aldehyde dehydrogenase
Biochem. J.
397
131-138
2006
Thermoplasma acidophilum (Q9HK01), Thermoplasma acidophilum, Thermoplasma acidophilum JCM 9062 (Q9HK01)
Manually annotated by BRENDA team
Reher, M.; Schnheit, P.
Glyceraldehyde dehydrogenases from the thermoacidophilic euryarchaeota Picrophilus torridus and Thermoplasma acidophilum, key enzymes of the non-phosphorylative Entner-Doudoroff pathway, constitute a novel enzyme family within the aldehyde dehydrogenase superfamily
FEBS Lett.
580
1198-1204
2006
Picrophilus torridus (Q6L285), Picrophilus torridus, Picrophilus torridus DSM 9790 (Q6L285), Thermoplasma acidophilum (Q9HK01), Thermoplasma acidophilum, Thermoplasma acidophilum JCM 9062 (Q9HK01)
Manually annotated by BRENDA team
Steffler, F.; Sieber, V.
Refolding of a thermostable glyceraldehyde dehydrogenase for application in synthetic cascade biomanufacturing
PLoS One
8
e70592
2013
Thermoplasma acidophilum (Q9HK01), Thermoplasma acidophilum, Thermoplasma acidophilum JCM 9062 (Q9HK01)
Manually annotated by BRENDA team
Steffler, F.; Guterl, J.K.; Sieber, V.
Improvement of thermostable aldehyde dehydrogenase by directed evolution for application in Synthetic Cascade Biomanufacturing
Enzyme Microb. Technol.
53
307-314
2013
Thermoplasma acidophilum (Q9HK01)
Manually annotated by BRENDA team
Iermak, I.; Degtjarik, O.; Steffler, F.; Sieber, V.; Kuta Smatanova, I.
Crystallization behaviour of glyceraldehyde dehydrogenase from Thermoplasma acidophilum
Acta Crystallogr. Sect. F
71
1475-1480
2015
Thermoplasma acidophilum (Q9HK01)
Manually annotated by BRENDA team
Iermak, I.; Degtjarik, O.; Steffler, F.; Sieber, V.; Kuta Smatanova, I.
Crystallization behaviour of glyceraldehyde dehydrogenase from Thermoplasma acidophilum
Acta Crystallogr. F Struct. Biol. Commun.
71
1475-1480
2015
Thermoplasma acidophilum (Q9HK01), Thermoplasma acidophilum, Thermoplasma acidophilum ATCC 25905 (Q9HK01)
Manually annotated by BRENDA team