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Literature summary for 3.1.1.11 extracted from
- Molecular cloning, expression and characterization of pectin methylesterase (CtPME) from Clostridium thermocellum (2017), Mol. Biotechnol., 59, 128-140 .
Application
| Application | Comment | Organism |
|---|---|---|
| industry | CtPME can be potentially used in food and textile industry applications | Acetivibrio thermocellus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene Cthe_2949, DNA and amino acid sequence determination and analysis and tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Acetivibrio thermocellus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Al3+ | - |
Acetivibrio thermocellus |  |
| Cu2+ | - |
Acetivibrio thermocellus | |
| EDTA | - |
Acetivibrio thermocellus | |
| EGTA | - |
Acetivibrio thermocellus | |
| Fe2+ | - |
Acetivibrio thermocellus | |
| Hg2+ | almost complete inhibition at 5 mM | Acetivibrio thermocellus | |
| additional information | no inhibition by Tween 20 and Triton X100 at 5 mM | Acetivibrio thermocellus | |
| SDS | - |
Acetivibrio thermocellus | |
| Tween 80 | - |
Acetivibrio thermocellus | |
| Urea | - |
Acetivibrio thermocellus | |
| Zn2+ | strong inhibition at 5 mM | Acetivibrio thermocellus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.1 | - |
Pectin | Citrus pectin (85% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 3.8 | - |
Pectin | apple pectin, recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 4.2 | - |
Pectin | Citrus pectin (75% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 4.7 | - |
Pectin | Citrus pectin (50% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 8.9 | - |
Pectin | Citrus pectin (25% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | 40% activation at 5 mM | Acetivibrio thermocellus | |
| Co2+ | activates at 5 mM | Acetivibrio thermocellus | |
| Cs+ | activates slightly at 5 mM | Acetivibrio thermocellus | |
| Mg2+ | 40% activation at 5 mM | Acetivibrio thermocellus | |
| Mn2+ | activates at 5 mM | Acetivibrio thermocellus | |
| additional information | no or poor effect by 5 mM of Ba2+, Li+, K+, and Cs+ | Acetivibrio thermocellus | |
| NaCl | activates slightly at 5 mM | Acetivibrio thermocellus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| pectin + n H2O | Acetivibrio thermocellus | - |
n methanol + pectate | - |
? | |
| pectin + n H2O | Acetivibrio thermocellus ATCC 27405 | - |
n methanol + pectate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Acetivibrio thermocellus | A3DJL8 | - |
- |
| Acetivibrio thermocellus ATCC 27405 | A3DJL8 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and dialysis | Acetivibrio thermocellus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 8.5 | - |
purified recombinant enzyme, pH 8.5, 50°C, substrate Citrus pectin DM 25% | Acetivibrio thermocellus |
| 12.3 | - |
purified recombinant enzyme, pH 8.5, 50°C, substrate Citrus pectin DM 50% | Acetivibrio thermocellus |
| 14.1 | - |
purified recombinant enzyme, pH 8.5, 50°C, substrate apple pectin | Acetivibrio thermocellus |
| 14.8 | - |
purified recombinant enzyme, pH 8.5, 50°C, substrate Citrus pectin DM 75% | Acetivibrio thermocellus |
| 18.1 | - |
purified recombinant enzyme, pH 8.5, 50°C, substrate Citrus pectin DM 85% | Acetivibrio thermocellus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | substrate specificity of CtPME is analysed against various pectic substrates (1%, w/v), viz. Citrus pectin of varying degrees of methyl esterification (85, 75-50 and 25%), apple pectin and poly galacturonic acid (PGA) from Citrus fruit, pectic galactan from potato and lupin, rhamnogalacturonan from soybean (RGS) and potato (RGP), overview | Acetivibrio thermocellus | ? | - |
? | |
| additional information | substrate specificity of CtPME is analysed against various pectic substrates (1%, w/v), viz. Citrus pectin of varying degrees of methyl esterification (85, 75-50 and 25%), apple pectin and poly galacturonic acid (PGA) from Citrus fruit, pectic galactan from potato and lupin, rhamnogalacturonan from soybean (RGS) and potato (RGP), overview | Acetivibrio thermocellus ATCC 27405 | ? | - |
? | |
| pectin + n H2O | - |
Acetivibrio thermocellus | n methanol + pectate | - |
? | |
| pectin + n H2O | best substrate is Citrus pectin with 85% methyl esterification | Acetivibrio thermocellus | n methanol + pectate | - |
? | |
| pectin + n H2O | - |
Acetivibrio thermocellus ATCC 27405 | n methanol + pectate | - |
? | |
| pectin + n H2O | best substrate is Citrus pectin with 85% methyl esterification | Acetivibrio thermocellus ATCC 27405 | n methanol + pectate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 35558, recombinant His6-tagged enzyme, MALDI-TOF mass spectrometry | Acetivibrio thermocellus |
| More | peptide mass fingerprint is performed by MALDI-TOF MS in reflectron mode with small molecule molecular weight range (500-5000 Da) | Acetivibrio thermocellus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Cthe_2949 | - |
Acetivibrio thermocellus |
| CtPME | - |
Acetivibrio thermocellus |
| pectin methylesterase | - |
Acetivibrio thermocellus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
recombinant enzyme | Acetivibrio thermocellus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | 70 | recombinant enzyme, CtPME retains 80% activity | Acetivibrio thermocellus |
| 50 | 60 | recombinant enzyme, CtPME retains 90% activity in the temperature range | Acetivibrio thermocellus |
| 80 | 85 | the protein melting curve of CtPME gives a peak at 80°C. The peak is shifted to 85°C in the presence of 5 mM Ca2+ ions, and the addition of 5 mM EDTA shifts back the melting peak to 80°C | Acetivibrio thermocellus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.72 | - |
Pectin | Citrus pectin (25% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 2.62 | - |
Pectin | Citrus pectin (50% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 3.06 | - |
Pectin | Citrus pectin (75% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 3.68 | - |
Pectin | apple pectin, recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 3.77 | - |
Pectin | Citrus pectin (85% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | - |
recombinant enzyme | Acetivibrio thermocellus |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 8 | 9 | recombinant enzyme, CtPME retains 85% activity | Acetivibrio thermocellus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | thermostable pectin methylesterase (CtPME) from Clostridium thermocellum belongs to family 8 carbohydrate esterase (CE8) | Acetivibrio thermocellus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.2 | - |
Pectin | Citrus pectin (25% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 0.55 | - |
Pectin | Citrus pectin (50% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 0.72 | - |
Pectin | Citrus pectin (75% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 0.97 | - |
Pectin | apple pectin, recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
| 1.21 | - |
Pectin | Citrus pectin (85% esterified), recombinant enzyme, pH 8.5, 50°C | Acetivibrio thermocellus | |
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