Literature summary extracted from
Hallin, E.I.; Guo, K.; Akerlund, H.E.
Violaxanthin de-epoxidase disulphides and their role in activity and thermal stability (2015), Photosynth. Res., 124, 191-198 .
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
1.23.5.1 |
gene VDE1, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) in inclusion bodies |
Spinacia oleracea |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
1.23.5.1 |
C09S |
site-directed mutagenesisthe mutant shows slightly decreased activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C118S |
site-directed mutagenesis, almost inactive mutant |
Spinacia oleracea |
1.23.5.1 |
C14S |
site-directed mutagenesis, almost inactive mutant |
Spinacia oleracea |
1.23.5.1 |
C21S |
site-directed mutagenesis, almost inactive mutant |
Spinacia oleracea |
1.23.5.1 |
C248S |
site-directed mutagenesis, the mutant shows over 95% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C27S |
site-directed mutagenesis, the mutant shows about 70% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C33S |
site-directed mutagenesis, the mutant shows about 90% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C37S |
site-directed mutagenesis, the mutant shows about 50% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C46S |
site-directed mutagenesis, the mutant shows about 45% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C50S |
site-directed mutagenesis, the mutant shows 90% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C65S |
site-directed mutagenesis, the mutant shows 85% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C72S |
site-directed mutagenesis, the mutant shows over 95% reduced activity compared to the wild-type enzyme |
Spinacia oleracea |
1.23.5.1 |
C7S |
site-directed mutagenesis, the mutant shows 2fold increased activity compared to the wild-type enzyme |
Spinacia oleracea |
Localization
EC Number |
Localization |
Comment |
Organism |
GeneOntology No. |
Textmining |
---|
1.23.5.1 |
chloroplast |
- |
Spinacia oleracea |
9507 |
- |
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
1.23.5.1 |
violaxanthin + 2 L-ascorbate |
Spinacia oleracea |
overall reaction |
zeaxanthin + 2 L-dehydroascorbate + 2 H2O |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.23.5.1 |
Spinacia oleracea |
Q9SM43 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
1.23.5.1 |
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) solubilized from inclusion bodies |
Spinacia oleracea |
Renatured (Commentary)
EC Number |
Renatured (Comment) |
Organism |
---|
1.23.5.1 |
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) inclusion bodies by solibilization with 8 M urea, 60 mM Tris-HCl, pH 8.0, 60 mM NaCl, and 0.6 mM EDTA, followed by dialysis and ultracentrifugation |
Spinacia oleracea |
Source Tissue
EC Number |
Source Tissue |
Comment |
Organism |
Textmining |
---|
1.23.5.1 |
leaf |
- |
Spinacia oleracea |
- |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.23.5.1 |
violaxanthin + 2 L-ascorbate |
overall reaction |
Spinacia oleracea |
zeaxanthin + 2 L-dehydroascorbate + 2 H2O |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
1.23.5.1 |
More |
enzyme VDE consists of a cysteine-rich N-terminal domain, a lipocalin-like domain and a negatively charged C-terminal domain. A disulphide pattern in VDE of C9-C27, C14-C21, C33-C50, C37-C46, C65-C72 and C118-C284 is obtained after digestion of VDE with thermolysin followed by mass spectroscopy analysis of reduced versus non-reduced samples. Reduction of the disulfides results in loss of a rigid structure and a decrease in thermal stability of 15°C. Peptide mapping, mass spectroscopy, overview |
Spinacia oleracea |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
1.23.5.1 |
VDE |
- |
Spinacia oleracea |
Temperature Stability [°C]
EC Number |
Temperature Stability Minimum [°C] |
Temperature Stability Maximum [°C] |
Comment |
Organism |
---|
1.23.5.1 |
33 |
- |
apparent melting temperature for the VDE reduced by DTT |
Spinacia oleracea |
1.23.5.1 |
48 |
- |
apparent melting temperature for the non-reduced VDE |
Spinacia oleracea |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
1.23.5.1 |
L-ascorbate |
- |
Spinacia oleracea |
|
General Information
EC Number |
General Information |
Comment |
Organism |
---|
1.23.5.1 |
malfunction |
mutational reduction of the disulfides in the enzyme results in loss of a rigid structure and a decrease in thermal stability of 15°C |
Spinacia oleracea |
1.23.5.1 |
physiological function |
violaxanthin de-epoxidase (VDE) catalyses the conversion of violaxanthin to zeaxanthin at the lumen side of the thylakoids during exposure to intense light |
Spinacia oleracea |