Cloned (Comment) | Organism |
---|---|
multiple DQD/SDH protein sequences are obtained from Phytozome 12.0 and NCBI, phylogenetic analysis and tree, recombinant expression of CsDQD/SDH isozymes in Escherichia coli strain NovaBlue (DE3) | Camellia sinensis |
Protein Variants | Comment | Organism |
---|---|---|
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 | Camellia sinensis |
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 | Camellia sinensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.046 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with shikimate | Camellia sinensis | |
0.115 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with shikimate | Camellia sinensis | |
0.138 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with shikimate | Camellia sinensis | |
0.2 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with NADP+ | Camellia sinensis | |
0.249 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with 3-dehydroshikimate | Camellia sinensis | |
0.267 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with 3-dehydroshikimate | Camellia sinensis | |
0.27 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with 3-dehydroshikimate | Camellia sinensis | |
0.273 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with NADP+ | Camellia sinensis | |
0.287 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with NADPH | Camellia sinensis | |
0.331 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with NADPH | Camellia sinensis | |
0.466 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHd, with NADPH | Camellia sinensis | |
0.611 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with NADP+ | Camellia sinensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
shikimate + NADP+ | Camellia sinensis | - |
3-dehydroshikimate + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Camellia sinensis | A0A3G3BK08 | four isozymes CsDQD/SDHa-d | - |
Purification (Comment) | Organism |
---|---|
recombinant CsDQD/SDH isozymes from Escherichia coli strain NovaBlue (DE3) by maltose affinity chromatography and ultrafiltration | Camellia sinensis |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
bud | - |
Camellia sinensis | - |
leaf | first and second | Camellia sinensis | - |
additional information | activities and phenolic contents in different tissues from CsDQD/SDH isozymes a-d, overview | Camellia sinensis | - |
root | - |
Camellia sinensis | - |
stem | - |
Camellia sinensis | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
relative activities for 3-dehydrochikimate reduction and shikimate oxidation of purified recombinant isozymes, overview | Camellia sinensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
shikimate + NADP+ | - |
Camellia sinensis | 3-dehydroshikimate + NADPH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
3-dehydroquinate dehydratase/shikimate dehydrogenase | - |
Camellia sinensis |
CsDQD/SDH | - |
Camellia sinensis |
DQD/SDH | - |
Camellia sinensis |
More | cf. EC 4.2.1.10 | Camellia sinensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Camellia sinensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.006 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with shikimate | Camellia sinensis | |
0.043 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with shikimate | Camellia sinensis | |
0.046 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHd, with NADPH | Camellia sinensis | |
0.049 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with 3-dehydroshikimate | Camellia sinensis | |
0.052 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with 3-dehydroshikimate | Camellia sinensis | |
0.085 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with NADP+ | Camellia sinensis | |
0.107 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with NADP+ | Camellia sinensis | |
0.117 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with shikimate | Camellia sinensis | |
0.153 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with NADP+ | Camellia sinensis | |
0.168 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with NADPH | Camellia sinensis | |
0.202 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with 3-dehydroshikimate | Camellia sinensis | |
0.405 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with NADPH | Camellia sinensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Camellia sinensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Camellia sinensis | |
NADPH | - |
Camellia sinensis |
General Information | Comment | Organism |
---|---|---|
evolution | four CsDQD/SDH isozyme proteins are cloned from Camellia sinensis. Three CsDQD/SDH isozymes show 3-dehydroshikimate (3-DHS) reduction and shikimate (SA) oxidation functions with individual differences between the catalytic efficiency of 3-DHS reduction and SA oxidation. Isozyme CsDQD/SDHa has higher catalytic efficiency for 3-DHS reduction than for SA oxidation, isozyme CsDQD/SDHd shows the opposite tendency, and isozyme CsDQD/SDHc has almost equal catalytic efficiency for 3-DHS reduction and SA oxidation. In vitro, Gallic acid (GA) is mainly generated from 3-DHS through nonenzymatic conversion. Isozymes CsDQD/SDHc and CsDQD/ SDHd genes are involved in GA synthesis | Camellia sinensis |
malfunction | the mutation of residues Ser338 and NRT to Gly and DI/LD in the SDH unit is the reason for the low activity of isozyme CsDQD/SDHb for 3-DHS reduction and SA oxidation | Camellia sinensis |
metabolism | 3-dehydroquinate dehydratase/shikimate dehydrogenase (DQD/SDH) is a key enzyme for catalyzing the conversion of 3-dehydroshikimate (3-DHS) to shikimate (SA). It also potentially participates in gallic acid (GA) synthesis in a branch of the SA pathway. Gallic acid (GA) is a precursor for polyphenol synthesis. The CsDQD/SDHc and CsDQD/ SDHd genes are involved in GA synthesis. In plants, DQD/SDH, a bifunctional enzyme, is crucial in the third and fourth reversible reactions in the SA pathway. Biosynthetic pathway of gallic acid and shikimic acid in plants, overview. GA could be spontaneously generated from 3-DHS in the enzymatic or nonenzymatic CsDQD/SDHs assay when using 3-DHS and the coenzyme NADP+ as substrates | Camellia sinensis |
physiological function | 3-dehydroquinate dehydratase/shikimate dehydrogenase (DQD/SDH) is a key enzyme for catalyzing the conversion of 3-dehydroshikimate (3-DHS) to shikimate (SA). It also potentially participates in gallic acid (GA) synthesis in a branch of the SA pathway. Quantitative RT-PCR analysis shows that CsDQD/SDHc and CsDQD/SDHd expression is correlated with GA and 1-O-galloyl-beta-D-glucose accumulation in Camellia sinensis. The SA pathway is important to plant growth, development, and defense. Polyphenols in tea plants, including phenolic acids, catechins, and flavonol derivatives, not only determine the mouthfeel of tea infusions but also provide health benefits | Camellia sinensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.099 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHd, with NADPH | Camellia sinensis | |
0.13 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with shikimate | Camellia sinensis | |
0.139 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with NADP+ | Camellia sinensis | |
0.195 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with 3-dehydroshikimate | Camellia sinensis | |
0.197 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHD, with 3-dehydroshikimate | Camellia sinensis | |
0.374 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with shikimate | Camellia sinensis | |
0.508 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with NADPH | Camellia sinensis | |
0.535 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHc, with NADP+ | Camellia sinensis | |
0.56 | - |
shikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with NADP+ | Camellia sinensis | |
0.748 | - |
NADPH | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with 3-dehydroshikimate | Camellia sinensis | |
0.848 | - |
NADP+ | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with shikimate | Camellia sinensis | |
1.411 | - |
3-dehydroshikimate | pH 7.5, 30°C, recombinant isozyme CsDQD/SDHa, with NADPH | Camellia sinensis |