Information on EC 1.1.1.132 - GDP-mannose 6-dehydrogenase

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

EC NUMBER
COMMENTARY hide
1.1.1.132
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RECOMMENDED NAME
GeneOntology No.
GDP-mannose 6-dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
GDP-D-mannose + 2 NAD+ + H2O = GDP-D-mannuronate + 2 NADH + 2 H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
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redox reaction
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-
-
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reduction
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
alginate biosynthesis
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alginate biosynthesis I (algal)
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alginate biosynthesis II (bacterial)
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Amino sugar and nucleotide sugar metabolism
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Fructose and mannose metabolism
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SYSTEMATIC NAME
IUBMB Comments
GDP-D-mannose:NAD+ 6-oxidoreductase
Also acts on the corresponding deoxynucleoside diphosphate derivative as a substrate.
CAS REGISTRY NUMBER
COMMENTARY hide
37250-63-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Arthrobacter sp. NRRL B1973
NRRL B1973
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
deoxy-GDP-D-mannose + NAD+ + H2O
deoxy-GDP-D-mannuronate + NADH
show the reaction diagram
GDP-D-mannose + NAD+ + H2O
GDP-D-mannunorate + NADH
show the reaction diagram
GDP-D-mannose + NAD+ + H2O
GDP-D-mannuronate + NADH
show the reaction diagram
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
show the reaction diagram
additional information
?
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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
GDP-D-mannose + NAD+ + H2O
GDP-D-mannunorate + NADH
show the reaction diagram
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the enzyme catalyzes the first committed step in alginate biosynthesis
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?
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CoCl2
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at 1 mM CoCl2 44% enzymatic activity
CuSO4
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at 1 mM CuSO4 no enzymatic activity
FeCl3
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at 1 mM FeCl3 49% enzymatic activity
FeSO4
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at 1 mM FeSO4 77% enzymatic activity
K+
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increases activity
Na+
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increases activity
PCMB
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0.025 mM, 0.8% activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5'-(3-hydroxypropyl)-amino-5'-deoxy-guanosine
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only after preincubation with enzyme
5'-azido-5'-deoxy-guanosine
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only after preincubation with enzyme
5'-azido-5'-deoxy-N2-2',3'-O-triacetylguanosine
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only after preincubation with enzyme
5'[[[[(2'',3'',4'',6''-tetra-O-acetyl 6''-ethynyl)alpha-D-mannopyranosyl oxy]-carbonyl]amino]sulfonyl]-2',3'-isopropylidene N2-acetyl guanosine
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GDP-analog, 0.1 mM, 60% inhibition
ambroxol
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i.e. 2-amino-3,5-dibromo-N-[trans-4-hydroxycyclohexyl]benzylamine, inhibits GMD activity, reduces the production of alginate, the expression of the important genes, and affects the structure of mucoid Pseudomonas aeruginosa biofilms, overview; reduces activity of the key enzyme GMD in alginate biosynthesis in a concentration-dependent manner
ATP
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; 26% inhibition at 1 mM
D-maltose
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17% inhibition at 1 mM
erythromycin
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potent dose-dependent inhibitory effect, inhibition of biofilm formation, PT-1578
GDP-D-glucose
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; 25% inhibition at 1 mM
guanosine
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slight inhibition only after preincubation with enzyme
iodacetamide
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40% inactivation within 30 min, complete reactivation by dithiothreitol
iodoacetamide
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40% inactivation after 30 min preincubation
midecamycin
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weaker inhibition, no biofilm destruction, PT-1578
N2-acetyl-2',3'-isopropylidene guanosine
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only after preincubation with enzyme
N2-acetyl-5'-amino-5'-deoxy-2',3'-O-acetylguanosine
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only after preincubation with enzyme
N2-acetyl-5'-amino-5'-deoxy-guanosine
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only after preincubation with enzyme
N2-acetyl-guanosine
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only after preincubation with enzyme
NADPH
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weak inhibitor of reaction
p-hydroxymercuribenzoate
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96% inactivation after 5 min preincubation; 96% inactivation within 5 min, complete reactivation by dithiothreitol
penicillic acid
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irreversible inactivation
tetra-O-acetylmannopyranose
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GDP-analog, 0.1 mM, 75% inhibition
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additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.084
deoxy-GDP-mannose
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-
0.009 - 0.095
GDP-D-mannose
0.017
GDP-mannose
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0.086 - 0.51
NAD+
additional information
additional information
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in phosphate buffer, enzyme shows Michaelis-Menten kinetics, in Tris buffer, enzyme shows marked cooperativity with respect to NAD+ binding. Phosphate and GMP are allosteric effectors
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0227
GMP
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24C, pH 8.0
additional information
additional information
-
inactivation kinetics of recombinant wild-type and mutant enzymes with penicillic acid
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00015
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+/-0.00001, S17-1/pPT4, with IPTG
0.00017
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+/-0.00002, S17-1/pMMB66EH and S17-1/pPT4 without IPTG
0.00022
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+/-0.00001, S17-1/pMMB66EH, with IPTG
0.00034
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+/-0.00006, S17-1/pPT3, without IPTG
0.0004
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+/-0.00008, PAO1/pMMB66EH, mid-exponential, without IPTG
0.00056
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+/-0.00024, PAO1/pMMB66EH, early-stationary, with IPTG
0.00076
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+/-0.00016, FRD2/pMMB66EH, mid-exponential, with IPTG
0.00089
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+/-0.0002, FRD2/pMMB66EH, mid-exponential, without IPTG
0.0009
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+/-0.0002, PAO1/pMMB66EH, with IPTG; +/-0.0003, PAO1/pMMB66EH, without IPTG
0.00091
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+/-0.00042, PAO1/pMMB66EH, mid-exponential, with IPTG
0.00094
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+/-0.00052, FRD2/pMMB66EH, early-stationary, with IPTG
0.00106
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+/-0.00062, PAO1/pMMB66EH, early-stationary, without IPTG
0.0013
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+/-0.00004, FRD2/pMMB66EH, early-stationary, without IPTG
0.002
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inorganic phosphate limitation, 85 h after changing carbon source from sorbitol to sucrose
0.002 - 0.015
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mucoid forms, activity correlated with alginate production and level of transcription of algD gene
0.0025
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oxygen limitation
0.0058
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+/-0.002, 8821/pMMB24, not IPTG induced
0.00685
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+/-0.00228, FRD1/pMMB66EH, mid-exponential, with IPTG
0.00749
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+/-0.00187, PAO1/pPT3, mid-exponential, without IPTG
0.00936
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+/-0.00063, PAO1/pPT3, early-stationary, without IPTG
0.00968
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+/-0.00155, FRD1/pMMB66EH, mid-exponential, without IPTG
0.00992
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+/-0.00105, PAO568/pMMB66EH, mid-exponential, without IPTG
0.01097
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+/-0.00195, FRD2/pPT4, early-stationary, with IPTG
0.01232
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+/-0.00397, PAO568/pMMB66EH, mid-exponential, with IPTG
0.01249
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+/-0.00113, FRD2/pPT4, early-stationary, without IPTG
0.0128
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+/-0.0018, 8821/pVD211, not IPTG induced
0.01283
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+/-0.00348, FRD2/pPT3, early-stationary, without IPTG
0.013
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+/-0.003, PAO1/pPT3, without IPTG
0.01312
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+/-0.0031, FRD2/pPT3, mid-exponential, without IPTG
0.014
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+/-0.0014, PAO1/pPT4, with IPTG
0.01401
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+/-0.00147, PAO1/pPT4, early-stationary, with IPTG
0.01436
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+/-0.00303, FRD1/pPT3, mid-exponential, without IPTG
0.0147
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+/-0.0014, PAO1/pPT4, without IPTG
0.01472
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+/-0.00135, PAO1/pPT4, early-stationary, without IPTG
0.0176
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+/-0.0009, PAO568/pMMB66EH, with IPTG
0.0182
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+/-0.0003, PAO568/pMMB66EH, without IPTG
0.01886
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+/-0.00526, PAO568/pMMB66EH, early-stationary, without IPTG
0.01891
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+/-0.0001, PAO568/pPT3, mid-exponential, without IPTG
0.0223
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+/-0.0011, PAO568/pPT3, without IPTG
0.0226
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+/-0.0025, 8821/pVD211, induced by 0.5 mM IPTG
0.02322
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+/-0.00432, PAO568/pMMB66EH, early-stationary, with IPTG
0.02418
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+/-0.00628, FRD1/pMMB66EH, early-stationary, without IPTG
0.0251
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+/-0.0027, PAO568/pPT4, without IPTG
0.02514
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+/-0.00274, PAO568/pPT4, early-stationary, without IPTG
0.0254
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+/-0.004, PAO568/pPT4, with IPTG
0.02542
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+/-0.00139, PAO568/pPT4, early-stationary, with IPTG
0.02574
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+/-0.00581, FRD1/pMMB66EH, early-stationary, with IPTG
0.0265
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+/-0.003, 8821/pVD211, induced by 3 mM IPTG
0.02671
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+/-0.00459, FRD1/pPT4, early-stationary, with IPTG
0.028
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+/-0.0029, 8821M/pMMB24, not IPTG induced
0.02905
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+/-0.00427, FRD1/pPT4, early-stationary, without IPTG
0.02994
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+/-0.0066, PAO568/pPT3, early-stationary, without IPTG
0.03208
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+/-0.00629, FRD1/pPT3, early-stationary, without IPTG
0.0579
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+/-0.0029, 8821/pVD211, not IPTG induced
0.084
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+/-0.0019, 8821/pVD211, induced by 0.5 mM IPTG
0.0942
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+/-0.004, 8821/pVD211, induced by 3 mM IPTG
0.1259
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+/-0.02878, PAO1/pPT3, mid-exponential, with IPTG
0.1364
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+/-0.01055, FRD2/pPT3, mid-exponential, with IPTG
0.1627
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+/-0.03188, PAO568/pPT3, mid-exponential, with IPTG
0.1758
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+/-0.03971, FRD1/pPT3, mid-exponential, with IPTG
0.1895
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+/-0.00702, S17-1/pPT3, with IPTG
0.1919
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+/-0.00268, PAO568/pPT3, early-stationary, with IPTG
0.2037
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+/-0.06998, PAO1/pPT3, early-stationary, with IPTG
0.2041
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+/-0.00932, PAO1/pPT3, with IPTG
0.2097
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+/-0.03274, FRD1/pPT3, early-stationary, with IPTG
0.2196
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+/-0.05517, FRD2/pPT3, early-stationary, with IPTG
0.2378
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+/-0.0552, PAO568/pPT3, with IPTG
1068
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24C, pH 8.0
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.2
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Tris buffer
8.75
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.75 - 9
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optimal pH range, enzyme inactive below pH 7
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34000
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SDS-PAGE
47600
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estimation of open reading frame
70000
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gel filtration
290000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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2 * 35000, gel filtration
hexamer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ternary complex with NAD and product GDP-mannuronic acid
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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enzyme partly inactivated above 30C
45
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enzyme fully inactivated at 45C after 15 min
50
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above, 2-3 min, loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
repeated freezing/thawing leads to decrease of activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, protein concentration above 1 mg/ml, at least 1 month
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-70C, 1-3 mg/ml purified protein, 50 mM triethanolamine acetate, pH 7.6, 1 mM DTT
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate fractionation, affinity chromatography
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ammonium sulfate precipitation
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pH/heat treatment acetone precipitation HPLC gel filtration chromatography
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protamine sulfate fractionation
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protamine sulfate precipitation, gel exclusion, ion exchange chromatography
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recombinant wild-type and mutant enzymes from Escherichia coli by gel filtration of enzyme with tightly bound GDP-mannuronic acid and NAD+, dialysis and ion exchange chromatography
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using Ni-NTA chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
algD cloned into a pMD18 T-vector and transformed into Escherichia coli DH5alpha-competent cells
expressed in Escherichia coli as a His-tagged fusion protein
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gene algD, overexpression of wild-type and mutant enzymes in Escherichia coli
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
in Azotobacter vinelandii GacA regulates alginate synthesis through its control of rsmZ1 and rsmZ2 transcription, which in turn relieves the repressing activity of RsmA on algD expression
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C213A
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site-directed mutagenesis, 1.8fold increased Vmax compared to the wild-type enzyme
C268A
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site-directed mutagenesis, 250fold reduced Vmax, 5fold increased KM, and reduced sensitivity to penicillic acid inactivation compared to the wild-type enzyme
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
inactivation by iodacetamide or p-hydroxymercuribenzoate is completely reversible
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