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Literature summary for 3.1.1.14 extracted from
- Development of a high-throughput purification method and a continuous assay system for chlorophyllase (2006), Anal. Biochem., 353, 93-98.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression as His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Triticum aestivum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-hydroxymercuribenzoate | - |
Triticum aestivum |  |
| Cu2+ | - |
Triticum aestivum | |
| diisopropyl fluorophosphate | - |
Triticum aestivum | |
| Fe2+ | - |
Triticum aestivum | |
| Fe3+ | - |
Triticum aestivum | |
| Hg2+ | - |
Triticum aestivum | |
| iodoacetamide | - |
Triticum aestivum | |
| Mg2+ | - |
Triticum aestivum | |
| additional information | inhibitory effects on recombinant enzymes purified by microtiter plate method and by affinity chromatography, respectively, overview, no inhibition by iodoacetic acid, iodoacetamide, 2-mercaptoethanol, and DTT | Triticum aestivum | |
| N-ethylmaleimide | - |
Triticum aestivum | |
| phenylmethanesulfonyl fluoride | - |
Triticum aestivum | |
| Zn2+ | - |
Triticum aestivum | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | steady state kinetics, the recombinant immobilized fusion protein displays kinetic parameters similar to those of recombinant enzyme purified by affnity chromatography, overview | Triticum aestivum | |
| 0.05 | - |
4-nitrophenyl decanoate | pH 8.0, 25°C, recombinant enzyme purified by affinity chromatography | Triticum aestivum | |
| 0.069 | - |
4-nitrophenyl decanoate | pH 8.0, 25°C, recombinant enzyme purified by microtiter plate method | Triticum aestivum | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| chlorophyll + H2O | Triticum aestivum | chlorophyllase catalyzes the initial hydrolysis of the phytol moiety from the pigment in the degradation of chlorophyll, overview | phytol + chlorophyllide | - |
? | |
| additional information | Triticum aestivum | since chlorophyll degradation is a defining feature of plant senescence, compounds inhibiting chlorophyllase activity may delay senescence, thereby improving shelf life and appearance of plant products | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Triticum aestivum | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, or by using a microtiter plate purification method with immobilization of the enzyme, overview | Triticum aestivum |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
development of a continuous assay system, replacing chlorophyll with p-nitrophenyl-ester substrates eliminates the extraction step and allows for continuous measurement of chlorophyllase activity in the multiwell plate format, overview | Triticum aestivum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl butyrate + H2O | - |
Triticum aestivum | 4-nitrophenol + butyrate | - |
? | |
| 4-nitrophenyl decanoate + H2O | - |
Triticum aestivum | 4-nitrophenol + decanoate | - |
? | |
| chlorophyll + H2O | - |
Triticum aestivum | phytol + chlorophyllide | - |
? | |
| chlorophyll + H2O | chlorophyllase catalyzes the initial hydrolysis of the phytol moiety from the pigment in the degradation of chlorophyll, overview | Triticum aestivum | phytol + chlorophyllide | - |
? | |
| additional information | since chlorophyll degradation is a defining feature of plant senescence, compounds inhibiting chlorophyllase activity may delay senescence, thereby improving shelf life and appearance of plant products | Triticum aestivum | ? | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Triticum aestivum |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 8.05 | - |
4-nitrophenyl decanoate | pH 8.0, 25°C, recombinant enzyme purified by affinity chromatography | Triticum aestivum | |
| 9.43 | - |
4-nitrophenyl decanoate | pH 8.0, 25°C, recombinant enzyme purified by microtiter plate method | Triticum aestivum | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at, substrate chlorophyll | Triticum aestivum |
| 8 | - |
assay at, substrate 4-nitrophenyl esters | Triticum aestivum |
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