Information on EC 1.1.1.6 - glycerol dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.1.1.6
-
RECOMMENDED NAME
GeneOntology No.
glycerol dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
glycerol + NAD+ = glycerone + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
glycerol degradation II
-
-
glycerol degradation V
-
-
Glycerolipid metabolism
-
-
Propanoate metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
glycerol:NAD+ 2-oxidoreductase
Also acts on propane-1,2-diol.
CAS REGISTRY NUMBER
COMMENTARY hide
249285-11-8
-
9028-14-2
this number also refers to EC 1.1.99.22
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
isolated from antarctic soil
-
-
Manually annotated by BRENDA team
mutant D
-
-
Manually annotated by BRENDA team
mutant D
-
-
Manually annotated by BRENDA team
DSM 2343
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2R,3R)-2,3-butanediol + NAD+
(3R)-acetoin + NADH
show the reaction diagram
(2R,3R)-2,3-butanediol + NAD+
? + NADH
show the reaction diagram
(R)-1,2-propanediol + NAD+
? + NADH
show the reaction diagram
(R)-1,2-propanediol + NAD+
hydroxyacetone + NADH
show the reaction diagram
1,2,3-butanetriol + NAD+
1,3-dihydroxybutane-2-one + NADH
show the reaction diagram
-
-
-
?
1,2-butanediol + NAD+
1-hydroxybutane-2-one + NADH
show the reaction diagram
-
-
-
r
1,2-butanediol + NAD+
? + NADH
show the reaction diagram
1,2-propanediol + NAD+
hydroxyacetone + NADH
show the reaction diagram
1,3-butanediol + NAD+
4-hydroxy-2-butanone + NADH + H+
show the reaction diagram
1,3-butanediol + NAD+
?
show the reaction diagram
-
relative activity is 6.5% compared to oxidation of glycerol
-
-
?
1,3-dichloro-2-propanol + NAD+
1,3-dichloro-2-propanone + NADH
show the reaction diagram
1,3-propanediol + NAD+
?
show the reaction diagram
1,4-butanediol + NAD+
?
show the reaction diagram
1-chloro-2,3-propanediol + NAD+
?
show the reaction diagram
1-phenylethan-1,2-diol + NAD+
? + NADH
show the reaction diagram
-
-
-
-
?
2,3-butanediol + NAD+
3-hydroxybutane-2-one + NADH
show the reaction diagram
3-amino-1,2-propanediol + NAD+
?
show the reaction diagram
-
104% of the activity with glycerol
-
-
?
3-bromo-1,2-propanediol + NAD+
?
show the reaction diagram
-
109% of the activity with glycerol
-
-
?
3-chloro-1,2-propanediol + NAD+
?
show the reaction diagram
-
130% of the activity with glycerol
-
-
?
3-hydroxypropionaldehyde + NADH
propan-1,3-diol + NAD+
show the reaction diagram
-
-
-
-
?
3-mercapto-1,2-propanediol + NAD+
?
show the reaction diagram
-
155% of the activity with glycerol
-
-
?
acetaldehyde + NADH + H+
ethanol + NAD+
show the reaction diagram
acetoin + NADH + H+
2,3-butanediol + NAD+
show the reaction diagram
beta-glycerophosphate + NAD+
?
show the reaction diagram
-
relative activity is 2% compared to oxidation of glycerol
-
-
?
butane-1,3-diol + NAD+
? + NADH
show the reaction diagram
-
-
-
-
?
diglycerol + NAD+
?
show the reaction diagram
-
relative activity is 21% compared to oxidation of glycerol
-
-
?
DL-alpha-glycerophosphate + NAD+
?
show the reaction diagram
DL-glyceraldehyde + NAD+
3-hydroxypyruvaldehyde + NADH
show the reaction diagram
DL-glyceraldehyde + NADH
glycerol + NAD+
show the reaction diagram
erythrite + NAD+
?
show the reaction diagram
-
-
-
-
?
ethanediol + NAD+
glycolaldehyde + NADH
show the reaction diagram
ethanol + NAD+
acetaldehyde + NADH
show the reaction diagram
-
relative activity is 1% compared to oxidation of glycerol
-
-
?
ethylene glycol + NAD+
?
show the reaction diagram
glycerol + NAD+
D-glyceraldehyde + NADH
show the reaction diagram
glycerol + NAD+
dihydroxyacetone + NADH
show the reaction diagram
glycerol + NAD+
dihydroxyacetone + NADH + H+
show the reaction diagram
glycerol + NAD+
glycerone + NADH
show the reaction diagram
-
-
-
-
?
glycerol + NAD+
glycerone + NADH + H+
show the reaction diagram
glycerol + NADP+
glyceraldehyde + NADPH + H+
show the reaction diagram
glycerol-alpha-monochlorohydrin + NAD+
?
show the reaction diagram
glycerol-alpha-monomethyl ether + NAD+
?
show the reaction diagram
glycerone + NADH + H+
glycerol + NAD+
show the reaction diagram
hydroxy-2-propanone + NADH
propylene glycol + NAD+
show the reaction diagram
-
relative activity is 27% compared to oxidation of glycerol
-
-
?
i-inositol + NAD+
?
show the reaction diagram
-
relative activity is 18% compared to oxidation of glycerol
-
-
?
isopropanol + NAD+
acetone + NADH
show the reaction diagram
-
relative activity is 17% compared to oxidation of glycerol
-
-
?
meso-2,3-butanediol + NAD+
(3S)-acetoin + NADH
show the reaction diagram
methylglyoxal + NADH
lactaldehyde + NAD+
show the reaction diagram
N-butyraldehyde + NADH
1-butanol + NAD+
show the reaction diagram
-
-
-
-
?
propane-1,2-diol + NAD+
propane-1-ol-2-one + NADH
show the reaction diagram
-
-
-
-
?
propionaldehyde + NADH
1-propanol + NAD+
show the reaction diagram
-
-
-
-
?
R-1-amino-2-propanol + NAD+
?
show the reaction diagram
-
33% of the activity with glycerol
-
-
?
S-1-amino-2-propanol + NAD+
?
show the reaction diagram
-
9% of the activity with glycerol
-
-
?
sorbitol + NAD+
?
show the reaction diagram
-
relative activity is 3% compared to oxidation of glycerol
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
glycerol + NAD+
D-glyceraldehyde + NADH
show the reaction diagram
glycerol + NAD+
dihydroxyacetone + NADH
show the reaction diagram
glycerol + NAD+
glycerone + NADH + H+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
N6-carboxymethyl-NAD+
-
wild-type enzyme TmGlyDH shows little activity with N6-carboxymethyl-NAD+ (N6-CM-NAD), an NAD+ analogue modified for easy immobilization to amino groups, but the double mutation V44A/K157G increases catalytic efficiency with N6-CMNAD+ by 10fold. kinetics with N6-CM-NAD+ and modelling, overview. N6-CM-NAD+ is immobilized on cyanogen bromide-activated Sepharose 4b
-
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
1 mM, stimulates 64.5% compared to 1 mM Mn2+
Co2+
-
presence of Mn2+ enhances the enzyme activity by 31.4%
Cu2+
-
presence of Mn2+ enhances the enzyme activity by 37.0%
Fe2+
-
1 mM, stimulates 26.3% compared to 1 mM Mn2+
Mg2+
improvement of stability, activity, and substrate promiscuity of glycerol dehydrogenase substituted by divalent metal ions Mn2+ and Mg2+, overview. The activity of Mn-GDH and Mg-GDH improves several folds in comparison to the native GDH. The activity of substituted GDH towards non-natural substrates, 4-chloroacetoacetate, 3-chloroacetylpyridine, p-chloroacetophenone, and acetophenone is 30folds higher than native GDH. Manganese substitution increases the half-life of GDH by 6folds at 60C and 70C
Na+
-
oxidation and reduction slightly increased
Tl+
-
activates
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
1,3-Propanediol
-
competitive inhibitor versus glycerol in direction of glycerol oxidation, noncompetitive inhibitor versus NAD+, competitive inhibitor versus dihydroxyacetone in direction of glycerol reduction, noncompetitive inhibitor versus NADH
1-ethyl-3(3-dimethylaminopropyl)carbodiimide
-
-
2,2'-bipyridyl
2,2'dipyridyl
-
1 mM, complete inhibition
2,4,6-Trinitrobenzene sulfonate
-
-
2-amino-2-(hydroxymethyl)-1,3-propanediol
-
strong inhibition, IC50 2 mM
2-mercaptoethanol
3,4-dimercaptotoluol
-
-
8-hydroxyquinoline
-
-
8-Quinolinol
acetone
-
inactivation, enzyme regains activity after removal of ketone
ADP
-
3 mM, 10% inhibition
AMP
-
3 mM, 13% inhibition
Anhitol 24B
-
-
ATP
-
3 mM, 6% inhibition
catechol
-
i.e. 1,2-butanediol, inactivation, enzyme regains activity after removal of alcohol
Cd2+
-
strongly inhibitory
Cetylpyridinium chloride
-
strong inhibition at low concentration
Cetyltrimethylammonium bromide
-
strong inhibition at low concentration
CuCl2
-
strong
D-xylose
-
-
diacetyl
-
inactivation, enzyme regains activity after removal of ketone
diethyldithiocarbamate
-
slight inhibition, 2 mM, 10 mM
dihydroxyacetone
dihydroxyacetone phosphate
-
-
dimethylformamide
-
inactivation
Dimethylsulfoxide
-
inactivation
dioxane
-
inactivation, enzyme regains activity after removal of ketone
diphosphate
-
-
dithioerythritol
-
10 mM
dithiothreitol
ethanol
-
10% v/v: 25-75% loss of activity, depending on substrate, enzyme regains activity after removal of alcohol
ethylene glycol
-
strong inhibition, IC50 4 mM
FeCl3
-
strong
glycerol 3-phosphate
-
-
HgCl2
-
strong
Hydroxyacetone
-
-
iodoacetamide
-
strong inhibition, 10 mM
iodoacetate
-
-
isobutanol
-
5% v/v: 0-50% loss of activity, depending on substrate, enzyme regains activity after removal of alcohol
K+
-
reduction of dihydroxyacetone
L-cysteine
Li+
-
competitive, forward reaction
meso-erythritol
-
-
methanol
-
10% v/v: 25-75% loss of activity, 5% v/v, 50-95% loss of activity, depending on substrate, enzyme regains activity after removal of alcohol
methylene blue
-
enzyme activity is decreased by photooxidation
Mg2+
-
inhibits the enzyme at high concentrations
n-amyl alcohol
-
2% v/v: 0-50% loss of activity, depending on substrate, enzyme regains activity after removal of alcohol
n-butanol
-
5% v/v: 0-50% loss of activity, depending on substrate, enzyme regains activity after removal of alcohol
N-ethylmaleimide
n-Propanol
-
5% v/v: 25-60% loss of activity, depending on substrate, enzyme regains activity after removal of alcohol
Na+
-
competitive, forward reaction
NADP+
Ni2+
-
inhibits the enzyme at high concentrations
o-phthalaldehyde
-
inactivation due to intramolecular thioisoindole formation, glycerol partially protects
p-chloromercuribenzoate
p-hydroxymercuribenzoate
-
0.1 mM, strong
Phenanthroline
-
1 mM, complete inhibition
polyethyleneimines
-
activating at 0.5 mM, inhibitory at 1 mM
-
pyridin-2,6-dicarboxylic acid
-
-
pyridoxyl-5'-phosphate
-
inactivation, NAD+ or NADH protect
Sodium dodecyl sulfate
-
-
sodium tetraborate
-
1 mM, 53% inhibition
Sucrose
-
-
additional information
-
enzyme inhibition simulations, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
-
at 1.85 mM 135% of the activity without activator
2-mercaptoethanol
-
1 mM, slight activation
8-Quinolinol
-
1 mM, slight activation
ammonium sulfate
-
about 100% activation at 10 mM
citrate
-
the enzyme is more active in presence of multivalent anions, citrate, phosphate or sulfate, in comparison to monovalent anions, e.g. acetate or chloride
dithioerythritol
-
1 mM, slight activation
iodoacetamide
-
at 1 mM 140% of the activity without activator, at 10 mM 220% of the activity without activator
K+
-
activates NADH production
N-ethylmaleimide
-
at 0.1 mM 145% of the activity without activator
p-chloromercuribenzoic acid
-
at 0.1 mM 133% of the activity without activator
phosphate
-
the enzyme is more active in presence of multivalent anions, citrate, phosphate or sulfate, in comparison to monovalent anions, e.g. acetate or chloride
polyethyleneimines
-
PEI, activating at 0.5 mM, inhibitory at 1 mM
-
SO42-
-
the enzyme is more active in presence of multivalent anions, citrate, phosphate or sulfate, in comparison to monovalent anions, e.g. acetate or chloride
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.06 - 0.202
1,2-propanediol
11.9 - 77.8
1,3-butanediol
0.0238
2,3-Butanediol
-
-
180
3-amino-1,2-propanediol
-
pH 8.8, 25C
6.1
3-bromo-1,2-propanediol
-
pH 8.8, 25C
6
3-Chloro-1,2-propanediol
-
pH 8.8, 25C
4
3-mercapto-1,2-propanediol
-
pH 8.8, 25C
0.06 - 4.87
dihydroxyacetone
10.9
glycero
-
-
-
0.5 - 118
glycerol
0.15 - 1.08
Glycerone
0.08 - 2.9
N6-CM-NAD+
0.0165 - 4.7
NAD+
0.014 - 0.12
NADH
4.4
R-1-amino-2-propanol
-
pH 8.8, 25C
500
S-1-amino-2-propanol
-
pH 8.8, 25C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.6 - 87.2
1,3-butanediol
1.89 - 3.36
glycerol
98.44
Glycerone
-
pH 6.0, 60C, recombinant enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.09 - 1.26
1,3-butanediol
0.06 - 129
glycerol
91.15
Glycerone
-
pH 6.0, 60C, recombinant enzyme
46.1 - 500
NAD+
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.1
dihydroxyacetone
-
-
11.6
Hydroxyacetone
-
-
1.09
NAD+
-
25C
0.025
NADH
-
25C
additional information
additional information
-
preliminary kinetic parameters of substrate and product inhibition, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2
2-amino-2-(hydroxymethyl)-1,3-propanediol
Pichia membranifaciens
-
strong inhibition, IC50 2 mM
0.3
2-mercaptoethanol
Pichia membranifaciens
-
strong inhibition, IC50 0.3 mM
4
ethylene glycol
Pichia membranifaciens
-
strong inhibition, IC50 4 mM
0.2
N-ethylmaleimide
Pichia membranifaciens
-
rapid inactivation, IC50: 0.2 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.01
wild type, crude cell extract, pH not specified in the publication, temperature not specified in the publication
0.033
-
anaerobic growth conditions, cells permeabilized with chloroform
0.049
overexpressing cells, crude cell extract, pH not specified in the publication, temperature not specified in the publication
1.05
-
cell free extract
1.26
-
; pH 8.2, 25C
3.7
-
pH 9.5, 30C
3.9
-
pH 9.5, 30C
4.7
-
cell free extract, wild type; wild type, crude extract, pH not specified in the publication, temperature not specified in the publication
4.8
-
wild type protein, pH 9.5, 30C
6
-
mutant protein D16N/N19A/E23D/L28M/E30N/R31N/Q45E/S46E/V48L/E49R/F52L/K53T/D54G/V58S/G78V/I79V/T82K/A83S/I88V/G108N/R139S/L142M/N145R/K155Q/L211I/G248S/V256I/H268Y/D317E/P319L, pH 9.5, 30C
6.8
-
mutant protein A15T/D16G/V17A/N19K/E23D/Q45E/S46E/T47M/V48L/E49R/F52L/K53A/V58A/V59A/Q70H/D74N/G78D/E81G/T82N/Q83K/G86T/I88V/G108N/R139S/L142M/N145R/K155Q/V256I/L260M, pH 9.5, 30C
7.4
-
mutant protein Q70H/D74N/G78D/E81G/T82N/Q83K/C84Y/G86T/I88V/G108N/E134A/E204K/L211I/E215K/I234V/V256I/L260M/E291D/S300C/A302S/E316G/V318I/A320T/I324L/T344D/P345S, pH 9.5, 30C
13.2
-
mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30C
15.7
-
recombinant His-tagged chimeric enzyme GDH-NOX, pH 11.0, 37C
45
-
purified recombinant enzyme, oxidation of glycerol, 80C, pH 7.4
90.7
-
mutant protein D121A, pH 9.5, 30C
107
-
commercial preparation
126.6
-
mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30C
220
-
cell free extract, recombinant strain (over-expression of gycerol dehydrogenase); recombinant strain (overexpression), crude extract, pH not specified in the publication, temperature not specified in the publication
additional information
-
assay development for NADH production by recombinant enzyme in reverse micelles, micelle size optimization, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
reduction of hydroxyacetone
5 - 6
-
reduction of dihydroxyacetone
7.2
-
oxidation of glycerol, wild-type enzyme
7.5
-
oxidation of glycerol, mutant D
7.5 - 8.5
-
reduction of dihydroxyacetone, phosphate buffer
8 - 8.5
-
oxidation of 1,2-propanediol
8.3
-
reduction of dihydroxyacetone, mutant D
8.6
-
reduction of dihydroxyacetone, wild-type enzyme
9.5 - 10
-
-
9.5 - 10.5
-
oxidation of glycerol, diphosphate buffer
10
-
oxidation of glycerol
10 - 11
-
the immobilized GDH changes the optimal pH from pH 11 to 10 and exhibits high activity in a relatively wide range of pH from pH 7 to 12, while free GDH only achieves high activity at pH 11
10.5 - 11
-
NADH production
10.5
-
in reverse micelles
11 - 12
-
oxidation of glycerol
additional information
-
kinetic study of the metal ion-chelated polyethyleneimines-immobilized enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 9
-
dihydroxyaceton reduction reaction, activity range, profile overview
4.8 - 7.8
-
pH 4.8: about 40% of maximal activity, pH 7.8: about 50% of maximal activity, reduction of dihydroxyacetone
5.5 - 9
-
pH 5.5: about 70% of maximal activity, pH 9.0: about 45% of maximal activity, reduction of dihydroxyacetone
6 - 11
6 - 8.5
-
oxidation activity range
6.4 - 9
-
reduction activity range
6.5 - 11
-
-
6.8 - 11
-
the initial rate of reaction increases upon increasing the pH from pH 6.8 to 11.0, past pH 11.0 the initial rate falls rapidly, oxidation of glycerol
7 - 10
7.5 - 12
-
glycerol oxidation reaction, activity range, profile overview
8 - 10.5
-
pH 8.0: about 40% of maximal activity, pH 10.5: about 70% of maximal activity, oxidation of glycerol
9 - 11
-
pH 9.0: maximal activity, pH 11.0.: nearly inactive, oxidation of glycerol
9 - 12
-
pH 9.0: about 45% of maximal activity, pH 11-12: maximal activity, oxidation of glycerol
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11
-
recombinant His-tagged chimeric enzyme GDH-NOX
25
-
assay at, purified enzyme
37
-
assay at, in reverse micelles
65
-
oxidation of propanediol
80
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
43% residual activity
20 - 80
-
activity range, profile overview
20 - 70
-
temperature profile, overview
90
-
reduction activity
additional information
-
kinetic study of the metal ion-chelated polyethyleneimines-immobilized enzyme
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
-
2 * 36000, SDS-PAGE
37000
-
-
38742
x * 38742 , calculated, 367 amino acid residues
39800
-
8 * 40000, recombinant His-tagged enzyme, SDS-PAGE, 8 * 39800, about, sequence calculation
42000
-
x * 42000, SDS-PAGE
47000
-
8 * 47000, SDS-PAGE
72000
-
gel filtration
76000 - 80000
-
gel filtration, PAGE under nondenaturing conditions
132000
-
gel filtration
156000 - 160000
-
density gradient centrifugation, gel filtration
160000
-
recombinant His6-tagged enzyme, ge filtration
180000
-
gel filtration
181000
-
gel filtration
320000
-
recombinant His-tagged enzyme, gel filtration
324000
-
gel filtration
336000 - 390000
400000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
octamer
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal obtained by hanging-drop vaour-diffusion method, S305C mutant
-
preliminary X-ray characterization; sitting drop vapor-diffusion method
hanging drop vapour diffusion, mixing of 0.001 ml 6 mg/ml protein in 150 mM ammonium acetate, 50 mM sodium chloride, 20 mM Tris, pH 7.4, 5% glycerol with 0.001 ml reservoir solution containing 5-10% PEG 3350, 0.2 M calcium acetate, and 4% glycerol, X-ray diffraction structure determination and analysis at 1.90 A resolution, molecular replacement using PDB entry 1jpu as a search model
-
in presence of glycerol
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
25C, 10 min, 30% loss of activity
636294
5.5 - 10
-
30C, 60 min, stable
636289
10
-
25C, 10 min, 40% loss of activity
636294
additional information
-
immobilization of the enzyme by metal ion-chelated polyethyleneimines greatly broadens the pH adaptability of GDH from strong alkaline into alkalescence
741417
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
60 min, stable in the pH range pH 5.5-10
30 - 50
purified recombinant enzyme, pH 12.0, quite stable over 150 min
55
-
recombinant enzyme, maintaining 70% activity after 2 h
60 - 70
-
recombinant enzyme, maintaining over 80% activity after 30 min
60
-
recombinant enzyme, maintaining 65% activity after 2 h
75
-
10 min stable
80
-
the recombinant enzyme, 40% activity remaining after 25 min, 20% activity remaining after 150 min
85
-
metal ion-chelated polyethyleneimine-immobilized enzyme PEI-Mn2+-GDH exhibits a half-life that is enhanced by 5.6folds in aqueous phase at 85C
additional information
thermal inactivation of GDH is not a simple first-order process, thermal deactivation kinetics, modelling, overview
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
manganese substitution increases the half-life of GDH by 6folds at 60C and 70C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, with 5% culture filtrate or 5% boiled crude cell extract, 10% Ficoll, pH 7.5, stable for 1 month
-
-70C, long term storage
-
0-5C, 50 to 55% loss of original activity within 48 h
-
4C, in 2 M (NH4)2SO4, 1 M glycerol, several months
-
frozen, retains about 42% of initial activity for 2 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene gldA, recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and dialysis
-
GST-tagged protein
-
immobilized metal ion affinity chromatography (Co2+)
immobilized metal ion affinity chromatography (Ni2+); purity estimated from SDS-PAGE: 95%
purified by Ni-affinity chromatography; recombinant His-tagged enzyme from strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged chimeric enzyme GDH-NOX from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by immobilized metal affinity chromatography or by heat shock treatment, both resulting in about the same protein purity of over 90%
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant mutant enzyme S305C
-
recombinant N-terminally His6-tagged enzyme from Escherichia coli train BL21(DE3)pLysS by nicke affinity chromatography and gel filtration
-
recombinent His-tagged enzyme from Escherichia coli strain BL21(DE3) pET-32a by nickel affinity chromatography, desalting gel filtration, and ultrafiltration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CglD, GST-tagged protein, expressed in E. coli BL21 (DE3)
-
cloning from contaminating bacteria, DNA and amino acid sequence determmination and analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli train BL21(DE3)pLysS
-
coexpression of enzymes recombinant glycerol dehydrogenase from Escherichia coli, lactate dehydrogenase form Bacillus subtilis, and NADH oxidase LpNox1 from Lactobacillus pentosus in Escherichia coli strain BL21(DE3). The activities of LDH, LpNox1 and GlyDH in the cell-free extract are 98.8, 27.8 and 39.5 U/ml, respectively, overview
-
expressed in E. coli
-
expressed in Klebsiella pneumoniae KG1
-
expressed in Klebsiella pneumoniae KG1; overexpressed in Klebsiella pneumoniae KG1
-
expression as His-tag fusion protein in Escherichia coli BL21 (DE3) pLysS; gene gld, expression of His-tagged enzyme in strain BL21(DE3)
-
expression in Saccharomyces cerevisiae
-
gene gldA from Escherichia coli strain JM109, expression of His-tagged enzyme in Escherichia coli strain BL21 (DE3), subcloning in Escherichia coli TOP10 cells
-
gene gldA, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain BL21(DE3)
-
gene gldA, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
gene gldA, recombinant expression in Escherichia coli strain BL21(DE3); gene gldA, recombinant expression in Escherichia coli strain BL21(DE3)
-
gene gldA, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
-
His-tag, expressed in E. coli JM109; His-tagged version
-
His-tagged version expressed in Escherichia coli BL21 STAR (DE3); N-terminal His-tag, expressed in E. coli BL21 STAR (DE3)
His-tagged version expressed in Escherichia coli BL21(DE3)
overexpressed in Schizosaccharomyces pombe
recombinant co-overexpression of enzymes glycerol dehydrogenase, malate dehydrogenase, and fumarate hydratase from Klebsiella pneumoniae subsp. pneumoniae strain ATCC 12657 in Propionibacterium jensenii leads to increased increased propionic acid production, quantittive reverse transcription PCR expression analysis
-
recombinant expression of the His-tagged enzyme encoding gene fused to the codon-optimized NADH oxidase gene nox from Lactobacillus brevis, chimeric enzyme GDH-NOX, in Escherichia coli strain BL21(DE3) resulting in bioconversion of glycerol to dihydroxyacetone in the cells
-
recombinent expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) pET-32a
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
induced by alcohols and aliphatic or aromatic hydrocarbons when glycerol used as the only substrate; induced by n-decanol (as sole carbon source)
induced by glucose starvation, essential for glycerol assimilation
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A15T/D16G/V17A/N19K/E23D/Q45E/S46E/T47M/V48L/E49R/F52L/K53A/V58A/V59A/Q70H/D74N/G78D/E81G/T82N/Q83K/G86T/I88V/G108N/R139S/L142M/N145R/K155Q/V256I/L260M
-
mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae)
D121A
-
D121 can potentially hinder the proper binding of substrate 1,3-butanediol due to steric hindrance
D16N/N19A/E23D/L28M/E30N/R31N/Q45E/S46E/V48L/E49R/F52L/K53T/D54G/V58S/G78V/I79V/T82K/A83S/I88V/G108N/R139S/L142M/N145R/K155Q/L211I/G248S/V256I/H268Y/D317E/P319L
-
mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae)
Q70H/D121A/G193C/E291Q/A310T
-
mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae) and site-directed mutation D121A
Q70H/D74N/G78D/E81G/T82N/Q83K/C84Y/G86T/I88V/G108N/E134A/E204K/L211I/E215K/I234V/V256I/L260M/E291D/S300C/A302S/E316G/V318I/A320T/I324L/T344D/P345S
-
mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae)
Q70H/G193C/E291Q/A310T
-
mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae)
S305C
-
S305C mutant used for crystallisation
I154A
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
I154A/K157G
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
I154A/K157N
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
K157G
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
K157N
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
V44A
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
V44A/I154A
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
V44A/K157G
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme; site-directed mutagenesis, wild-type enzyme TmGlyDH shows little activity with N6-carboxymethyl-NAD+ (N6-CM-NAD), an NAD+ analogue modified for easy immobilization to amino groups, but the double mutation V44A/K157G increases catalytic efficiency with N6-CMNAD+ by 10fold
V44A/K157N
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
I154A
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
-
K157N
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
-
V44A
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme
-
V44A/K157G
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
-
site-directed mutagenesis, the mutant shows altered activity with cofactor derivative N6-CM-NAD+ immobilized on Sepharose beads compared to the wild-type enzyme; site-directed mutagenesis, wild-type enzyme TmGlyDH shows little activity with N6-carboxymethyl-NAD+ (N6-CM-NAD), an NAD+ analogue modified for easy immobilization to amino groups, but the double mutation V44A/K157G increases catalytic efficiency with N6-CMNAD+ by 10fold
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
brewing
molecular biology
synthesis
Show AA Sequence (1940 entries)
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