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Literature summary for 3.1.1.26 extracted from
- A novel galactolipase from a green microalga Chlorella kessleri purification, characterization, molecular cloning, and heterologous expression (2018), Appl. Microbiol. Biotechnol., 102, 1711-1723 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| Triton X-100 | strongly activating, best at 0.3% w/v. Absence of this detergent causes the inactivation of ckGL even in the enzyme assay, as it does during the purification | Parachlorella kessleri |  |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene chGL or glp1, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21, subcloning in Escherichia coli strain JM109 | Parachlorella kessleri |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | - |
Parachlorella kessleri | |
| DTT | - |
Parachlorella kessleri | |
| EDTA | strong inhibition | Parachlorella kessleri | |
| Fe2+ | strong inhibition, complete inhibition at 10 mM | Parachlorella kessleri | |
| Fe3+ | strong inhibition, 98% inhibition at 10 mM | Parachlorella kessleri | |
| iodoacetamide | - |
Parachlorella kessleri | |
| PMSF | - |
Parachlorella kessleri | |
| Zn2+ | strong inhibition, complete inhibition at 10 mM | Parachlorella kessleri | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Parachlorella kessleri | |
| 0.54 | - |
1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol | pH 6.5, 37°C, native enzyme | Parachlorella kessleri | |
| 1.05 | - |
1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol | pH 6.5, 37°C, recombinant enzyme | Parachlorella kessleri | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| chloroplast | - |
Parachlorella kessleri | 9507 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | dependent on, best at 2 mM | Parachlorella kessleri | |
| Mn2+ | moderate activation | Parachlorella kessleri | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 146000 | 242000 | native enzyme, gel filtration and native PAGE | Parachlorella kessleri |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1,2-diacyl-3-beta-D-galactosyl-sn-glycerol + H2O | Parachlorella kessleri | - |
? | - |
? | |
| 1,2-diacyl-3-beta-D-galactosyl-sn-glycerol + H2O | Parachlorella kessleri 11h | - |
? | - |
? | |
| 1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol + H2O | Parachlorella kessleri | - |
? | - |
? | |
| 1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol + H2O | Parachlorella kessleri 11h | - |
? | - |
? | |
| additional information | Parachlorella kessleri | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol (DGDG) | ? | - |
? | |
| additional information | Parachlorella kessleri 11h | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol (DGDG) | ? | - |
? | |
| phosphatidylglycerol + H2O | Parachlorella kessleri | - |
? | - |
? | |
| phosphatidylglycerol + H2O | Parachlorella kessleri 11h | - |
? | - |
? | |
| sulfoquinovosyl diglyceride + H2O | Parachlorella kessleri | - |
? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Parachlorella kessleri | A0A2Z5TTR4 | i.e. Parachlorella kessleri | - |
| Parachlorella kessleri 11h | A0A2Z5TTR4 | i.e. Parachlorella kessleri | - |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| proteolytic modification | Thr61 is the predominant N-terminal amino acid of mature ckGL, Ala60 might be removed by an aminopeptidase after the transport of ckGL into the chloroplast | Parachlorella kessleri |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native enzyme 163fold from Chlorella kessleri strain 11h to homogeneity, it is not possible to purify active enzyme in the absence of Triton X-100 or by using other detergents because the enzyme becomes inactive during the procedure. Recombinant His-tagged enzyme 15.5fold from Escherichia coli strain JM109 by 2 steps of nickel affinity chromatography | Parachlorella kessleri |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 143 | - |
purified native enzyme, pH 6.5, 37°C | Parachlorella kessleri |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1,2-diacyl-3-beta-D-galactosyl-sn-glycerol + H2O | - |
Parachlorella kessleri | ? | - |
? | |
| 1,2-diacyl-3-beta-D-galactosyl-sn-glycerol + H2O | - |
Parachlorella kessleri 11h | ? | - |
? | |
| 1,2-diacyl-3-beta-D-galactosyl-sn-glycerol + H2O | - |
Parachlorella kessleri | 2-acyl-3-beta-D-galactosyl-sn-glycerol + fatty acid | - |
? | |
| 1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol + H2O | - |
Parachlorella kessleri | ? | - |
? | |
| 1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol + H2O | - |
Parachlorella kessleri 11h | ? | - |
? | |
| 1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol + H2O | - |
Parachlorella kessleri | 2-acyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol + fatty acid | - |
? | |
| 1,2-dimyristoyl-sn-glycero-3-phosphate + H2O | - |
Parachlorella kessleri | 2-myristoyl-sn-glycero-3-phosphate + myristate | - |
? | |
| 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-)glycerol + H2O | - |
Parachlorella kessleri | 2-oleoyl-sn-glycero-3-phospho-(1'-rac-)glycerol + palmitate | - |
? | |
| 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine + H2O | low activity | Parachlorella kessleri | 2-oleoyl-sn-glycero-3-phosphocholine + palmitate | - |
? | |
| 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine + H2O | low activity | Parachlorella kessleri | ? | - |
? | |
| additional information | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol (DGDG) | Parachlorella kessleri | ? | - |
? | |
| additional information | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol, substrate specificity in descending order: digalactosyldiacylglycerol (100%), monogalactosyldiacylglycerol and phosphatidylglycerol (about 40%), sulfoquinovosyldiacylglycerol (about 20%). The enzyme exhibits mostly no activity toward glycerides and other phospholipids. e.g. 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine and 1,2-dimyristoyl-sn-glycero-3-phosphate, or tristearin, diplamitin, and olive oil. ckGL preferably hydrolyzes the sn-1 acyl ester bond in the substrates | Parachlorella kessleri | ? | - |
? | |
| additional information | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol (DGDG) | Parachlorella kessleri 11h | ? | - |
? | |
| additional information | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol, substrate specificity in descending order: digalactosyldiacylglycerol (100%), monogalactosyldiacylglycerol and phosphatidylglycerol (about 40%), sulfoquinovosyldiacylglycerol (about 20%). The enzyme exhibits mostly no activity toward glycerides and other phospholipids. e.g. 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine and 1,2-dimyristoyl-sn-glycero-3-phosphate, or tristearin, diplamitin, and olive oil. ckGL preferably hydrolyzes the sn-1 acyl ester bond in the substrates | Parachlorella kessleri 11h | ? | - |
? | |
| phosphatidylglycerol + H2O | - |
Parachlorella kessleri | ? | - |
? | |
| phosphatidylglycerol + H2O | - |
Parachlorella kessleri 11h | ? | - |
? | |
| sulfoquinovosyl diglyceride + H2O | - |
Parachlorella kessleri | ? | - |
? | |
| sulfoquinovosyl diglyceride + H2O | lower activity | Parachlorella kessleri | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 53000, SDS-PAGE | Parachlorella kessleri |
| More | peptide sequencing | Parachlorella kessleri |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| chGL | - |
Parachlorella kessleri |
| ckGL | - |
Parachlorella kessleri |
| galactolipase | - |
Parachlorella kessleri |
| glp1 | - |
Parachlorella kessleri |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
- |
Parachlorella kessleri |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | 40 | purified native enzyme, pH 6.5, 30 min, completely stable at | Parachlorella kessleri |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 498 | - |
1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol | pH 6.5, 37°C, recombinant enzyme | Parachlorella kessleri | |
| 1390 | - |
1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol | pH 6.5, 37°C, native enzyme | Parachlorella kessleri | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
- |
Parachlorella kessleri |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 7 | purified native enzyme, 4°C, 30 min, completely stable at | Parachlorella kessleri |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Parachlorella kessleri | sequence calculation | - |
6.32 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | differences between galactolipases and lipases with regard to function, substrate recognition mechanism, and biochemical roles, overview. Phylogenetic analysis based on amino acid sequences also show that ckGL is clearly separated from groups containing phospholipase A1, PLA1 | Parachlorella kessleri |
| physiological function | galactolipase (ckGL) hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol (DGDG) | Parachlorella kessleri |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 474.3 | - |
1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol | pH 6.5, 37°C, recombinant enzyme | Parachlorella kessleri | |
| 2574.1 | - |
1,2-diacyl-3-O-alpha-D-galactosyl-(1->6)-beta-D-galactosyl-sn-glycerol | pH 6.5, 37°C, native enzyme | Parachlorella kessleri | |
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