Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
D423A
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, inactive mutant
D423N
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme
H335A
-
site-directed mutagenesis, tenfold decrease in kcat value
K385A
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme
K385N
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme
K385N/D423N
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme
N406A
-
site-directed mutagenesis, very strong decrease in kcat value
Q578L
-
site-directed mutagenesis, sixtyfold decrease in kcat value
Q582L
-
site-directed mutagenesis, very strong decrease in kcat value
S336A
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme
S338A
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme
S338G
site-directed mutagenesis, the mutant is not active with quinate like the wild-type
S338G/T381G
site-directed mutagenesis, the double mutant does not show improved enzymatic activity with quinate compared with the T381G mutant
T381G
site-directed mutagenesis, mutant shows increased activity with quinate compared to wild-type, it catalyzes the oxidation of quinate with a turnover rate of 8.8/s and a KM of 3.33 mM
T407A
-
site-directed mutagenesis, 6.5fold decrease in kcat value, increase in Km-value
T422S
-
site-directed mutagenesis, tenfold decrease in kcat value
Y550A
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme
Y550F
-
site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme
G338S/G381T/D483N/L484R/D485T
site-directed mutagenesis, mutant MTCsDQD/SDHb has a similar reduction activity of 3-DHS and had six times higher oxidation activity of SA than wild-type isozyme CsDQD/SDHb, suggesting that the mutation of residues Ser338 and NRT to Gly and DI/LD in the SDH unit is the reason for the low activity of CsDQD/SDHb, respectively
A243G
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
D195E
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
N149D
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
N149D/V152F
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
S131A
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
S131A/L135A
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
S131A/L135A/N149D/V152F
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
S131A/N149D/V152F
site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme
D103X
-
site-directed mutagenesis of paralogue HI0607, inactive mutant
K67H
-
site-directed mutagenesis of paralogue HI0607, inactive mutant
K69A
-
site-directed mutagenesis, the mutant shows significantly reduced the catalytic efficiency compared to wild-type enzyme
Q237A
-
site-directed mutagenesis
Q237K
-
site-directed mutagenesis
Q237N
-
site-directed mutagenesis
Y210A
-
site-directed mutagenesis
Y210S
-
site-directed mutagenesis
A213L
site-directed mutagenesis, analysis of substrate and cofactor binding compared to wild-type enzyme
D105A
-
the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid
K69H
-
the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid
K69I
-
the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid
K69Q
-
the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid
S275G
-
site-directed mutagenesis, the mutant shows only slightly reduced maximum activity with shikimate compared with wild-type PoptrSDH1
S275G/T318G
-
site-directed mutagenesis, the double mutant is well expressed in Escherichia coli and shows bona fide QDH activity besides its original SDH activity, which is severely reduced. Although the Ser275Gly/Thr318Gly double mutant is clearly sufficient to confer gain of activity with quinate, its activity is lower than the QDH activities of PintaQDH and PoptrQDH2 activity
T318G
-
site-directed mutagenesis, the Thr318Gly mutant yields only a very small amount of enzyme when recombinantly expressed in Escherichia coli
Y211F
results in a remarkable reduction in enzyme activity, leads to a significant decrease in kcat (345fold) and a minor increase in the Km (3fold) for shikimate. Tyr211 may play a major role in the catalytic process and a minor role in the initial substrate binding
T381A
-
site-directed mutagenesis, more than twentyfold decrease in kcat value
T381A
site-directed mutagenesis, the mutant accepts quinate as a substrate but is much less efficient than the T381G variant
T381S
-
site-directed mutagenesis, fourfold decrease in kcat value, slight decrease in Km value
T381S
site-directed mutagenesis, the mutant accepts quinate as a substrate but is much less efficient than the T381G variant
D105N
-
the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid
D105N
site-directed mutagenesis, analysis of substrate and cofactor binding compared to wild-type enzyme
K69A
-
the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid. Best recombinant protein expression protocol of the K69A mutant in insoluble form is using Escherichia coli C41(DE3) strain, grown at 37°C for 24 h after induction with 1 mM IPTG, and cell disruption by sonication
K69A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme, overview
K69A
site-directed mutagenesis, analysis of substrate and cofactor binding compared to wild-type enzyme
additional information
-
construction of deletion variant DELTA88DHQ-SDH, thermal denaturation analysis, overview
additional information
absence of the T381 side chain creates sufficient space in the active site to accommodate the quinate C1-hydroxyl. The K385 and D423 catalytic dyad which interacts with C4-OH and participates in proton transfer during the reduction/oxidation of NADP+/NADPH is retained in the T381G mutant
additional information
referring to the model of Arabidopsis thaliana protein crystal structure, the amino acid residue sites of Gly338, Gly381, Asp483, Leu484, and Asp485 in CsDQD/SDHb are mutated to Ser338, Thr381, Asn483, Arg484, and Thr485, respectively, the mutant protein is named MTCsDQD/SDHb
additional information
-
referring to the model of Arabidopsis thaliana protein crystal structure, the amino acid residue sites of Gly338, Gly381, Asp483, Leu484, and Asp485 in CsDQD/SDHb are mutated to Ser338, Thr381, Asn483, Arg484, and Thr485, respectively, the mutant protein is named MTCsDQD/SDHb
additional information
steady state cytosolic free and whole cell NADPH/NADP ratio in different Saccharomycs cerevisiae strains, with or without recombinant expression of shikimate dehydrogenase from Escherichia coli, thermodynamics and kinetics, overview
additional information
invertion of the cofactor specificity from NADP+ to NAD+ on the Escherichia coli wild-type enzyme, effect of consensus mutations, overview. Mutant structure modeling
additional information
-
invertion of the cofactor specificity from NADP+ to NAD+ on the Escherichia coli wild-type enzyme, effect of consensus mutations, overview. Mutant structure modeling
additional information
ydiB-encoded enzyme knockout in Escherichia coli strain PB12
additional information
-
ydiB-encoded enzyme knockout in Escherichia coli strain PB12
-
additional information
-
cytosolic isoform cannot complement loss of the plastidial isoform
additional information
cytosolic isoform cannot complement loss of the plastidial isoform
additional information
cytosolic isoform cannot complement loss of the plastidial isoform
additional information
-
expression of endogenous DHD/SHD-1 is suppressed by RNAi in transgenic tobacco plants, the transgenic lines with less than 40% of wild-type activity display severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin, but accumulation of dehydroquinate and shikimate, possibly due to existence of a parallel extra-plastidic shikimate pathway into which dehydroquinate is diverted with a second gene DHD/SHD-2 in tobacco lacking a plastidic targeting sequence, the cytosolic shikimate synthesis cannot complement loss of the plastidial pathway, phenotype, overview
additional information
expression of endogenous DHD/SHD-1 is suppressed by RNAi in transgenic tobacco plants, the transgenic lines with less than 40% of wild-type activity display severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin, but accumulation of dehydroquinate and shikimate, possibly due to existence of a parallel extra-plastidic shikimate pathway into which dehydroquinate is diverted with a second gene DHD/SHD-2 in tobacco lacking a plastidic targeting sequence, the cytosolic shikimate synthesis cannot complement loss of the plastidial pathway, phenotype, overview
additional information
expression of endogenous DHD/SHD-1 is suppressed by RNAi in transgenic tobacco plants, the transgenic lines with less than 40% of wild-type activity display severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin, but accumulation of dehydroquinate and shikimate, possibly due to existence of a parallel extra-plastidic shikimate pathway into which dehydroquinate is diverted with a second gene DHD/SHD-2 in tobacco lacking a plastidic targeting sequence, the cytosolic shikimate synthesis cannot complement loss of the plastidial pathway, phenotype, overview
additional information
-
suppression of shikimate dehydrogenase activity by RNAi. Transgenic lines with less than 40% of wild-type activity display severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin. Dehydroquinate, the substrate of the enzyme, accumulates, and due to a second, cytosolic enzyme, the product, shikimate accumulates
additional information
suppression of shikimate dehydrogenase activity by RNAi. Transgenic lines with less than 40% of wild-type activity display severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin. Dehydroquinate, the substrate of the enzyme, accumulates, and due to a second, cytosolic enzyme, the product, shikimate accumulates
additional information
suppression of shikimate dehydrogenase activity by RNAi. Transgenic lines with less than 40% of wild-type activity display severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin. Dehydroquinate, the substrate of the enzyme, accumulates, and due to a second, cytosolic enzyme, the product, shikimate accumulates
additional information
the shikimate C1-carboxyl is formed by the phenol hydroxyl of a tyrosine. Substitution of this residue in Staphyococcus epidermidis SDH causes a substantial reduction in turnover rate
additional information
-
the shikimate C1-carboxyl is formed by the phenol hydroxyl of a tyrosine. Substitution of this residue in Staphyococcus epidermidis SDH causes a substantial reduction in turnover rate
-