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Deoxy-GDP-D-mannose + 2 NAD+ + H2O
deoxy-GDP-D-mannuronate + 2 NADH + 2 H+
deoxy-GDP-D-mannose + NAD+ + H2O
deoxy-GDP-D-mannuronate + NADH
GDP-D-mannose + 2 NAD+ + H2O
GDP-D-mannuronate + 2 NADH + 2 H+
GDP-D-mannose + NAD+ + H2O
GDP-D-mannunorate + NADH
GDP-D-mannose + NAD+ + H2O
GDP-D-mannuronate + NADH
GDP-D-mannose + NAD+ + H2O
GDP-D-mannuronate + NADH + H+
-
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
additional information
?
-
Deoxy-GDP-D-mannose + 2 NAD+ + H2O
deoxy-GDP-D-mannuronate + 2 NADH + 2 H+
-
-
-
-
r
Deoxy-GDP-D-mannose + 2 NAD+ + H2O
deoxy-GDP-D-mannuronate + 2 NADH + 2 H+
-
-
-
-
r
deoxy-GDP-D-mannose + NAD+ + H2O
deoxy-GDP-D-mannuronate + NADH
-
-
-
-
?
deoxy-GDP-D-mannose + NAD+ + H2O
deoxy-GDP-D-mannuronate + NADH
-
-
-
?
deoxy-GDP-D-mannose + NAD+ + H2O
deoxy-GDP-D-mannuronate + NADH
-
-
-
-
?
GDP-D-mannose + 2 NAD+ + H2O
GDP-D-mannuronate + 2 NADH + 2 H+
-
-
-
?
GDP-D-mannose + 2 NAD+ + H2O
GDP-D-mannuronate + 2 NADH + 2 H+
-
-
-
?
GDP-D-mannose + NAD+ + H2O
GDP-D-mannunorate + NADH
-
-
-
-
?
GDP-D-mannose + NAD+ + H2O
GDP-D-mannunorate + NADH
-
the enzyme catalyzes the first committed step in alginate biosynthesis
-
-
?
GDP-D-mannose + NAD+ + H2O
GDP-D-mannuronate + NADH
-
-
-
-
?
GDP-D-mannose + NAD+ + H2O
GDP-D-mannuronate + NADH
-
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
ir
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
water-soluble and insoluble polysaccharides are produced, containing glucuronic and mannuronic acid, glucose and rhamnose, GDP-D-mannose dehydrogenase activity detected in 30 h cultures and 36 h cultures containing only water-insoluble capsular or water-soluble polysaccharides, respectively
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
?
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
ir
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
ir
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
ir
GDP-mannose + NAD+ + H2O
GDP-mannuronate + NADH
-
-
-
ir
additional information
?
-
-
key enzyme in alginate biosynthesis
-
-
?
additional information
?
-
-
no substrate: D-mannose, UDP-D-mannose, UDP-D-glucose, TDP-D-glucose, CDP-D-glucose
-
-
?
additional information
?
-
-
GMD is the key enzyme in alginate biosynthesis. Biofilm-forming bacteria Pseudomonas aeruginosa is a common pathogen in mechanically ventilated newborns, which can cause life-threatening infections
-
-
?
additional information
?
-
GMD is the key enzyme in alginate biosynthesis. Biofilm-forming bacteria Pseudomonas aeruginosa is a common pathogen in mechanically ventilated newborns, which can cause life-threatening infections
-
-
?
additional information
?
-
algD is localized in an alginate biosynthesis cluster
-
-
?
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5'-(3-hydroxypropyl)-amino-5'-deoxy-guanosine
-
only after preincubation with enzyme
5'-azido-5'-deoxy-guanosine
-
only after preincubation with enzyme
5'-azido-5'-deoxy-N2-2',3'-O-triacetylguanosine
-
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
-
GDP-analog, 0.1 mM, 60% inhibition
ambroxol
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
-
26% inhibition at 1 mM
C6-amido-GDP-D-mannose
mixed mechanism of inhibition
-
Ca2+
-
inhibitor of isoform GMD2 with 71.86% residual activity at 1 mM
D-maltose
-
17% inhibition at 1 mM
erythromycin
-
potent dose-dependent inhibitory effect, inhibition of biofilm formation, PT-1578
GDP-D-glucose
-
25% inhibition at 1 mM
guanosine
-
slight inhibition only after preincubation with enzyme
iodacetamide
-
40% inactivation within 30 min, complete reactivation by dithiothreitol
iodoacetamide
-
40% inactivation after 30 min preincubation
midecamycin
-
weaker inhibition, no biofilm destruction, PT-1578
N2-acetyl-2',3'-isopropylidene guanosine
-
only after preincubation with enzyme
N2-acetyl-5'-amino-5'-deoxy-2',3'-O-acetylguanosine
-
only after preincubation with enzyme
N2-acetyl-5'-amino-5'-deoxy-guanosine
-
only after preincubation with enzyme
N2-acetyl-guanosine
-
only after preincubation with enzyme
NADPH
-
weak inhibitor of reaction
p-hydroxymercuribenzoate
-
96% inactivation after 5 min preincubation; 96% inactivation within 5 min, complete reactivation by dithiothreitol
penicillic acid
-
irreversible inactivation
tetra-O-acetylmannopyranose
-
GDP-analog, 0.1 mM, 75% inhibition
Zn2+
-
potent inhibitor with 0.75% residual activity at 1 mM for both isoforms GMD1 and GMD2
GMP
-
competitive, 52% inhibition at 1 mM; competitive, most potent
additional information
-
full reactivation by dithiothreitol (25 mM) after p-hydroxymercuribenzoate or iodoacetamide inactivation
-
additional information
-
strains which resist 0.005 mg/ml of tobramycin treated by 5'-(3-hydroxypropyl)-amino-5'-deoxy-guanosine, tetra-O-acetylmannopyranose and 5'[[[[(2'',3'',4'',6''-tetra-O-acetyl 6''-ethynyl)alpha-D-mannopyranosyl oxy]-carbonyl]amino]sulfonyl]-2',3'-isopropylidene N2-acetyl guanosine recover 20, 75 and 85% of their sensisivity towards tobramycin
-
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0.00015
-
+/-0.00001, S17-1/pPT4, with IPTG
0.00017
-
+/-0.00002, S17-1/pMMB66EH and S17-1/pPT4 without IPTG
0.00022
-
+/-0.00001, S17-1/pMMB66EH, with IPTG
0.00034
-
+/-0.00006, S17-1/pPT3, without IPTG
0.0004
-
+/-0.00008, PAO1/pMMB66EH, mid-exponential, without IPTG
0.00056
-
+/-0.00024, PAO1/pMMB66EH, early-stationary, with IPTG
0.00076
-
+/-0.00016, FRD2/pMMB66EH, mid-exponential, with IPTG
0.00089
-
+/-0.0002, FRD2/pMMB66EH, mid-exponential, without IPTG
0.00091
-
+/-0.00042, PAO1/pMMB66EH, mid-exponential, with IPTG
0.00094
-
+/-0.00052, FRD2/pMMB66EH, early-stationary, with IPTG
0.00106
-
+/-0.00062, PAO1/pMMB66EH, early-stationary, without IPTG
0.0013
-
+/-0.00004, FRD2/pMMB66EH, early-stationary, without IPTG
0.002
-
inorganic phosphate limitation, 85 h after changing carbon source from sorbitol to sucrose
0.002 - 0.015
-
mucoid forms, activity correlated with alginate production and level of transcription of algD gene
0.0025
-
oxygen limitation
0.0058
-
+/-0.002, 8821/pMMB24, not IPTG induced
0.00685
-
+/-0.00228, FRD1/pMMB66EH, mid-exponential, with IPTG
0.00749
-
+/-0.00187, PAO1/pPT3, mid-exponential, without IPTG
0.00936
-
+/-0.00063, PAO1/pPT3, early-stationary, without IPTG
0.00968
-
+/-0.00155, FRD1/pMMB66EH, mid-exponential, without IPTG
0.00992
-
+/-0.00105, PAO568/pMMB66EH, mid-exponential, without IPTG
0.01097
-
+/-0.00195, FRD2/pPT4, early-stationary, with IPTG
0.01232
-
+/-0.00397, PAO568/pMMB66EH, mid-exponential, with IPTG
0.01249
-
+/-0.00113, FRD2/pPT4, early-stationary, without IPTG
0.0128
-
+/-0.0018, 8821/pVD211, not IPTG induced
0.01283
-
+/-0.00348, FRD2/pPT3, early-stationary, without IPTG
0.013
-
+/-0.003, PAO1/pPT3, without IPTG
0.01312
-
+/-0.0031, FRD2/pPT3, mid-exponential, without IPTG
0.014
-
+/-0.0014, PAO1/pPT4, with IPTG
0.01401
-
+/-0.00147, PAO1/pPT4, early-stationary, with IPTG
0.01436
-
+/-0.00303, FRD1/pPT3, mid-exponential, without IPTG
0.0147
-
+/-0.0014, PAO1/pPT4, without IPTG
0.01472
-
+/-0.00135, PAO1/pPT4, early-stationary, without IPTG
0.0176
-
+/-0.0009, PAO568/pMMB66EH, with IPTG
0.0182
-
+/-0.0003, PAO568/pMMB66EH, without IPTG
0.01886
-
+/-0.00526, PAO568/pMMB66EH, early-stationary, without IPTG
0.01891
-
+/-0.0001, PAO568/pPT3, mid-exponential, without IPTG
0.0223
-
+/-0.0011, PAO568/pPT3, without IPTG
0.0226
-
+/-0.0025, 8821/pVD211, induced by 0.5 mM IPTG
0.02322
-
+/-0.00432, PAO568/pMMB66EH, early-stationary, with IPTG
0.02418
-
+/-0.00628, FRD1/pMMB66EH, early-stationary, without IPTG
0.0251
-
+/-0.0027, PAO568/pPT4, without IPTG
0.02514
-
+/-0.00274, PAO568/pPT4, early-stationary, without IPTG
0.0254
-
+/-0.004, PAO568/pPT4, with IPTG
0.02542
-
+/-0.00139, PAO568/pPT4, early-stationary, with IPTG
0.02574
-
+/-0.00581, FRD1/pMMB66EH, early-stationary, with IPTG
0.0265
-
+/-0.003, 8821/pVD211, induced by 3 mM IPTG
0.02671
-
+/-0.00459, FRD1/pPT4, early-stationary, with IPTG
0.028
-
+/-0.0029, 8821M/pMMB24, not IPTG induced
0.02905
-
+/-0.00427, FRD1/pPT4, early-stationary, without IPTG
0.02994
-
+/-0.0066, PAO568/pPT3, early-stationary, without IPTG
0.03208
-
+/-0.00629, FRD1/pPT3, early-stationary, without IPTG
0.0579
-
+/-0.0029, 8821/pVD211, not IPTG induced
0.084
-
+/-0.0019, 8821/pVD211, induced by 0.5 mM IPTG
0.0942
-
+/-0.004, 8821/pVD211, induced by 3 mM IPTG
0.1259
-
+/-0.02878, PAO1/pPT3, mid-exponential, with IPTG
0.1364
-
+/-0.01055, FRD2/pPT3, mid-exponential, with IPTG
0.1627
-
+/-0.03188, PAO568/pPT3, mid-exponential, with IPTG
0.1758
-
+/-0.03971, FRD1/pPT3, mid-exponential, with IPTG
0.1895
-
+/-0.00702, S17-1/pPT3, with IPTG
0.1919
-
+/-0.00268, PAO568/pPT3, early-stationary, with IPTG
0.2037
-
+/-0.06998, PAO1/pPT3, early-stationary, with IPTG
0.2041
-
+/-0.00932, PAO1/pPT3, with IPTG
0.2097
-
+/-0.03274, FRD1/pPT3, early-stationary, with IPTG
0.2196
-
+/-0.05517, FRD2/pPT3, early-stationary, with IPTG
0.2378
-
+/-0.0552, PAO568/pPT3, with IPTG
0.0009
-
+/-0.0003, PAO1/pMMB66EH, without IPTG
0.0009
-
+/-0.0002, PAO1/pMMB66EH, with IPTG
additional information
-
-
additional information
-
increase of activity has only limited effect on the stimulation of alginate synthesis, existance of other rate-limiting enzymatic step suggested
additional information
-
presence of 1 mM GDP-mannose, GDP-glucose, GMP, GDP, GTP or cGMP retards tryptic digest of enzyme and protects subsequent loss of enzyme activity, amino-terminal domain contains substrate and cofactor binding sites, carboxy-terminal domain is essential for catalytic activity
additional information
-
higher enzyme activities after IPTG induction only for pPT3 containing strains, low GDP-mannuronate production with pMMB66EH only detectable in mucoid strains, higher with pPT3 with increase in cells of mucoid and non-mucoid strains with IPTG, alginate formation occurs only in mucoid strains without significant increase with IPTG, later enzyme of alginate biosynthesis suggested to become rate-limiting
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NRRL B1973
-
-
brenda
-
-
-
brenda
-
UniProt
brenda
-
-
-
brenda
-
UniProt
brenda
-
-
-
brenda
NRRL B1973
-
-
brenda
-
-
-
brenda
NCIB 8789, water soluble polysaccharide producing strains E, W and O
-
-
brenda
SM52B, mutant of A. vinelandii NCIB 9068, alginate-prducing ability, no activity with sorbitol as sole carbon source during batch growth, enzyme not produced, loss of ability to synthesise alginate during prolonged continuous culture after 300 h for mutant and more than 1000 h for parent strain, ability transferable by conjugation
-
-
brenda
-
-
-
brenda
-
UniProt
brenda
-
-
-
brenda
-
UniProt
brenda
A22 alg+, selected from mucoid strain IPA22
-
-
brenda
algD gene overproduced in strain 8822 using a plasmid vector containing algD under control of the tac promoter
-
-
brenda
enzyme overproduced using 3 different plasmid vectors containing algD under control of the tac promoter expressed in non-mucoid strain PAO1 and mucoid strain PAO568, pPT3 and pPT4 are forward and reverse clones contain leader sequence with possible transcriptional/translational regulatory sequences of algD gene, no significant activity with pMMB66EH and pPT4 in either IPTG induced or non induced Escherichia coli, mucoid and non-mucoid Pseudomonas aeruginosa strains
-
-
brenda
enzyme overproduced using a plasmid vector containing algD under control of the tac promoter
-
-
brenda
expressed in several mutants
-
-
brenda
gene algD
-
-
brenda
mucoid clinical isolate PT-1578 and non-mucoid PAO1, very low activity of PAO1 regardless whether using nutrient-rich or poor medium, low activity with PT-1578 on enriched, increased on nutritionally poor medium
-
-
brenda
mucoid strain 8821
-
-
brenda
mucoid strain PA7563
-
-
brenda
mucoid strains of cystic fibrosis patients 8821 and its high-alginate-producing spontaneously variant 8821M, enzyme overproduced using 2 different plasmid vectors containing algD under control of the tac promoter mobilized into both strains
-
-
brenda
mucoid strains of cystic fibrosis patients 8821, its high-alginate-producing spontaneously variant 8821M, IST23, IST27 and IST30, mucoid strains derived from the spontaneous nonmucoid strains during prolonged incubation under nutrient limitation in acetamide broth (Nm), rich in PolyP, no activity in spontaneous swiched nonmucoid variants (N) of initial isolated mucoid strains
-
-
brenda
no activity in nonmucoid mutants
-
-
brenda
overexpression of tac promoter controlled algD gene containing different plasmids in mucoid strains FRD1 and PAO568 and non-mucoid strains FRD2 and PAO1
-
-
brenda
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Preiss, J.
GDP-mannose dehydrogenase from Arthrobacter
Methods Enzymol.
8
285-287
1966
Arthrobacter sp., Arthrobacter sp. NRRL B1973
-
brenda
Roychoudhury, S.; Mmay, T.B.; Gill, J.F.; Singh, S.K.; Feingold, D.S.; Chakrabarty, A.M.
Purification and characterization of guanosine diphospho-D-mannose dehydrogenase. A key enzyme in the biosynthesis of alginate by Pseudomonas aeruginosa
J. Biol. Chem.
264
9380-9385
1989
Pseudomonas aeruginosa
brenda
Preiss, J.
Sugar nucleotide reactions in Arthrobacter. II. Biosynthesis of guanosine diphosphomannuronate
J. Biol. Chem.
239
3127-3132
1964
Arthrobacter sp.
brenda
Pugashetti, B.K.; Vadas, L.; Prihar, H.S.; Feingold, D.S.
GDPmannose dehydrogenase and biosynthesis of alginate-like polysaccharide in a mucoid strain of Pseudomonas aeruginosa
J. Bacteriol.
153
1107-1110
1983
Pseudomonas aeruginosa
brenda
Deretic, V.; Gill, J.F.; Chakrabarty, A.M.
Gene algD coding for GDPmannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa
J. Bacteriol.
169
351-358
1987
Pseudomonas aeruginosa
brenda
Horan, N.J.; Jarman, T.R.; Dawes, E.A.
Studies on some enzymes of alginic acid biosynthesis in Azotobacter vinelandii grown in continuous culture
J. Gen. Microbiol.
129
2985-2990
1983
Azotobacter vinelandii
-
brenda
Couperwhite, I.; McCallum, M.F.
Polysaccharide production and the possible occurrence of GDP-D-mannose dehydrogenase in Azotobacter vinelandii
Antonie van Leeuwenhoek
41
25-32
1975
Azotobacter vinelandii
brenda
Tavares, I.M.; Leitao, J.H.; Fialho, A.M.; Sa-Correia, I.
Pattern of changes in the activity of enzymes of GDP-D-mannuronic acid synthesis and in the level of transcription of algA, algC and algD genes accompanying the loss and emergence of mucoidy in Pseudomonas aeruginosa
Res. Microbiol.
150
105-116
1999
Pseudomonas aeruginosa
brenda
Martins, L.O.; Sa-Correia, I.
Alginate biosynthesis in mucoid recombinants of Pseudomonas aeruginosa overproducing GDP-mannose dehydrogenase
Enzyme Microb. Technol.
13
385-389
1991
Pseudomonas aeruginosa
brenda
Tatnell, P.J.; Russell, N.J.; Gacesa, P.
Overexpression of the GDP-mannose dehydrogenase gene from Pseudomonas aeruginosa
Biochem. Soc. Trans.
24
405S
1996
Pseudomonas aeruginosa
brenda
Elloumi, N.; Moreau, B.; Aguiar, L.; Jaziri, N.; Sauvage, M.; Hulen, C.; Capmau, M.L.
Inhibitors of GDP-mannose dehydrogenase of Pseudomonas aeruginosa mucoid strains
Eur. J. Med. Chem.
27
149-154
1992
Pseudomonas aeruginosa
-
brenda
Roychoudhury, S.; Chakrabarty, K.; Ho, Y.K.; Chakrabarty, A.M.
Characterization of guanosine diphospho-D-mannose dehydrogenase from Pseudomonas aeruginosa. Structural analysis by limited proteolyses
J. Biol. Chem.
267
990-996
1992
Pseudomonas aeruginosa
brenda
Mitsuya, Y.; Kawai, S.; Kobayashi, H.
Influence of macrolides on guanosine diphospho-D-mannose dehydrogenase activity in Pseudomonas biofilm
J. Infect. Chemother.
6
45-50
2000
Pseudomonas aeruginosa
brenda
Tatnell, P.J.; Russell, N.J.; Gacesa, P.
GDP-mannose dehydrogenase is the key regulatory enzyme in alginate biosynthesis in Pseudomonas aeruginosa: evidence from metabolite studies
Microbiology
140
1745-1754
1994
Pseudomonas aeruginosa
brenda
Naught, L.E.; Gilbert, S.; Imhoff, R.; Snook, C.; Beamer, L.; Tipton, P.
Allosterism and cooperativity in Pseudomonas aeruginosa GDP-mannose dehydrogenase
Biochemistry
41
9637-9645
2002
Pseudomonas aeruginosa
brenda
Snook, C.F.; Tipton, P.A.; Beamer, L.J.
Crystal structure of GDP-mannose dehydrogenase: a key enzyme of alginate biosynthesis in P. aeruginosa
Biochemistry
42
4658-4668
2003
Pseudomonas aeruginosa (P11759)
brenda
Kimmel, J.L.; Tipton, P.A.
Inactivation of GDP-mannose dehydrogenase from Pseudomonas aeruginosa by penicillic acid identifies a critical active site loop
Arch. Biochem. Biophys.
441
132-140
2005
Pseudomonas aeruginosa
brenda
Li, F.; Yu, J.; Yang, H.; Wan, Z.; Bai, D.
Effects of ambroxol on alginate of mature Pseudomonas aeruginosa biofilms
Curr. Microbiol.
57
1-7
2008
Pseudomonas aeruginosa, Pseudomonas aeruginosa (P11759)
brenda
Shi, X.; Gong, Q.; Cheng, R.
Detection of the key enzyme of alginate biosynthesis in Vibrio sp. QY102
World J. Microbiol. Biotechnol.
24
1613-1615
2008
Vibrio sp. QY102 (A4ZZ81)
brenda
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