Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosseta-Gami B (DE3) cells | Brassica oleracea |
Protein Variants | Comment | Organism |
---|---|---|
D170A | inactive | Brassica oleracea |
H226A | the mutation causes almost complete loss of Chlase activity | Brassica oleracea |
H239A | the mutation causes almost complete loss of Chlase activity | Brassica oleracea |
H247A | the mutation causes almost complete loss of Chlase activity | Brassica oleracea |
H66A | the mutation causes almost complete loss of Chlase activity | Brassica oleracea |
S141A | inactive | Brassica oleracea |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0764 | - |
pheophorbide a methyl ester | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.143 | - |
pheophorbide a methyl ester | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.308 | - |
chlorophyll a | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.311 | - |
chlorophyll b | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.466 | - |
chlorophyll a | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.652 | - |
chlorophyll b | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | isoform CLH2 | Brassica oleracea | 9507 | - |
chloroplast | isoforms CLH1 | Brassica oleracea | 9507 | - |
plasma membrane | isoform CLH3 | Brassica oleracea | 5886 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
23500 | - |
x * 23500, isoform CLH3, calculated from amino acid sequence | Brassica oleracea |
34700 | - |
x * 34700, isoform CLH1, calculated from amino acid sequence | Brassica oleracea |
35300 | - |
x * 35300, isoform CLH2, calculated from amino acid sequence | Brassica oleracea |
66200 | - |
x * 66200, isoform CLH3 as MBP-fusion protein, SDS-PAGE | Brassica oleracea |
77400 | - |
x * 77400, isoform CLH1 as MBP-fusion protein, SDS-PAGE | Brassica oleracea |
78000 | - |
x * 78000, isoform CLH2 as MBP-fusion protein, SDS-PAGE | Brassica oleracea |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Brassica oleracea | Q8GTM2 | isoform CLH3; cultivar Green king | - |
Brassica oleracea | Q8GTM3 | isoform CLH2; cultivar Green king | - |
Brassica oleracea | Q8GTM4 | isoform CLH1; cultivar Green king | - |
Purification (Comment) | Organism |
---|---|
amylose resin affinity column chromatography and DEAE Sepharose column chromatography | Brassica oleracea |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
floret | - |
Brassica oleracea | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.074 | - |
mutant enzyme H247A, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
0.149 | - |
mutant enzyme H239A, in 100mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
0.372 | - |
mutant enzyme H66A, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
0.52 | - |
mutant enzyme H226A, in 100mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
7.43 | - |
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
chlorophyll a + H2O | - |
Brassica oleracea | ? | - |
? | |
chlorophyll b + H2O | - |
Brassica oleracea | ? | - |
? | |
additional information | isoform CLH1 preferably hydrolyzes Mg-free chlorophyll | Brassica oleracea | ? | - |
? | |
additional information | isoform CLH2 hydrolyzes both chlorophyll and Mg-free chlorophyll at a similar level | Brassica oleracea | ? | - |
? | |
additional information | isoform CLH3 shows very low Chlase activity | Brassica oleracea | ? | - |
? | |
pheophorbide a methyl ester + H2O | - |
Brassica oleracea | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 23500, isoform CLH3, calculated from amino acid sequence | Brassica oleracea |
? | x * 34700, isoform CLH1, calculated from amino acid sequence | Brassica oleracea |
? | x * 35300, isoform CLH2, calculated from amino acid sequence | Brassica oleracea |
? | x * 66200, isoform CLH3 as MBP-fusion protein, SDS-PAGE | Brassica oleracea |
? | x * 77400, isoform CLH1 as MBP-fusion protein, SDS-PAGE | Brassica oleracea |
? | x * 78000, isoform CLH2 as MBP-fusion protein, SDS-PAGE | Brassica oleracea |
Synonyms | Comment | Organism |
---|---|---|
Chlase | - |
Brassica oleracea |
chlorophyll chlorophyllidohydrolase | - |
Brassica oleracea |
CLH1 | isoform | Brassica oleracea |
CLH2 | isoform | Brassica oleracea |
CLH3 | isoform | Brassica oleracea |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
isoform CLH1 | Brassica oleracea |
60 | - |
isoform CLH2 | Brassica oleracea |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
after incubation at 60°C for 10 min, isoform CLH1 loses about 50% of activity | Brassica oleracea |
60 | - |
after incubation at 60°C for 10 min, isoform CLH2 loses about 50% of activity | Brassica oleracea |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0024 | - |
chlorophyll a | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.0037 | - |
chlorophyll b | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.0039 | - |
pheophorbide a methyl ester | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.072 | - |
chlorophyll a | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.072 | - |
chlorophyll b | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.259 | - |
pheophorbide a methyl ester | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
isoform CLH1 | Brassica oleracea |
8 | - |
isoform CLH2 | Brassica oleracea |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | isoform CLH1 | Brassica oleracea |
8 | 10 | isoform CLH2 shows about 80% of activity at pH 9.0 and is stable between pH 8.0 and 10.0 | Brassica oleracea |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0078 | - |
chlorophyll a | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.0119 | - |
chlorophyll b | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.0272 | - |
pheophorbide a methyl ester | wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.1105 | - |
chlorophyll b | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
0.1551 | - |
chlorophyll a | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea | |
3.397 | - |
pheophorbide a methyl ester | wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40°C | Brassica oleracea |