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Literature summary for 3.2.1.157 extracted from
- Cloning, expression and characterization of a new iota-carrageenase from marine bacterium, Cellulophaga sp. (2013), Biotechnol. Lett., 35, 1617-1622.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene cgiB_Ce, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression in Escherichia coli strain BL21(DE3) | Cellulophaga sp. |
| gene cgiB_Ce, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Cellulophaga sp. |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Al3+ | 56% inhibition at 1 mM | Cellulophaga sp. |  |
| Cu2+ | 87% inhibition at 1 mM | Cellulophaga sp. | |
| EDTA | 90% inhibition at 1 mM | Cellulophaga sp. | |
| Fe3+ | 11% inhibition at 1 mM | Cellulophaga sp. | |
| SDS | 97% inhibition at 1 mM | Cellulophaga sp. | |
| Zn2+ | 49% inhibition at 1 mM | Cellulophaga sp. | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ba2+ | 118% activation at 1 mM | Cellulophaga sp. | |
| Ca2+ | 122% activation at 1 mM | Cellulophaga sp. | |
| Fe2+ | 174% activation at 1 mM | Cellulophaga sp. | |
| K+ | 134% activation at 10 mM | Cellulophaga sp. | |
| Li+ | 114% activation at 1 mM | Cellulophaga sp. | |
| Mg2+ | 149% activation at 1 mM | Cellulophaga sp. | |
| Mn2+ | 173% activation at 1 mM | Cellulophaga sp. | |
| Na+ | 166% activation at 500 mM | Cellulophaga sp. | |
| NaCl | although its activity is enhanced by NaCl, the enzyme is active in the absence of NaCl | Cellulophaga sp. | |
| Ni2+ | 128% activation at 1 mM | Cellulophaga sp. | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 48071 | - |
x * 48071, sequence calculation | Cellulophaga sp. |
| 48071 | - |
x * 48071, sequence calculation, x * 54000, recombinant His-tagged enzyme, SDS-PAGE | Cellulophaga sp. |
| 54000 | - |
x * 48071, sequence calculation, x * 54000, recombinant His-tagged enzyme, SDS-PAGE | Cellulophaga sp. |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| iota-carrageenan + H2O | Cellulophaga sp. | - |
neo-iota-carratetraose | - |
? | |
| iota-carrageenan + H2O | Cellulophaga sp. | yielding neo-iota-carratetraose as the main product in the absence of NaCl | neo-iota-carratetraose | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Cellulophaga sp. | R9UQG9 | gene cgiB_Ce | - |
| Cellulophaga sp. | R9UQG9 | isolated from the red alga Grateloupia livida collected from Qingdao coastal waters of China Yellow Sea, gene cgiB_Ce | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme from Escherichia coli strain BL21(DE3) culture supernatant by affinity chromatography | Cellulophaga sp. |
| recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Cellulophaga sp. |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 1870 | - |
pH 6.5, 45°C, with iota-carrageenan | Cellulophaga sp. |
| 1870 | - |
purified enzyme, pH 6.5, 45°C | Cellulophaga sp. |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| iota-carrageenan + H2O | - |
Cellulophaga sp. | neo-iota-carratetraose | - |
? | |
| iota-carrageenan + H2O | yielding neo-iota-carratetraose as the main product in the absence of NaCl | Cellulophaga sp. | neo-iota-carratetraose | - |
? | |
| iota-carrageenan + H2O | the endo-type iota-carrageenase hydrolyzes beta-1,4-linkages of iota-carrageenan | Cellulophaga sp. | neo-iota-carratetraose | neo-carratetraose is the main product with more than 80% of the total product | ? | |
| iota-carrageenan + H2O | the endo-type iota-carrageenase hydrolyzes beta-1,4-linkages of iota-carrageenan yielding neo-iota-carratetraose as the main product in the absence of NaCl, mass and NMR spectrometric product analysis, overview | Cellulophaga sp. | neo-iota-carratetraose | - |
? | |
| additional information | no activity towards jota-carrageenan, kappa-carrageenan or agarose | Cellulophaga sp. | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 48071, sequence calculation | Cellulophaga sp. |
| ? | x * 48071, sequence calculation, x * 54000, recombinant His-tagged enzyme, SDS-PAGE | Cellulophaga sp. |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CgiB_Ce | - |
Cellulophaga sp. |
| endo-type iota-carrageenase | - |
Cellulophaga sp. |
| iota-carrageenase | - |
Cellulophaga sp. |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | - |
- |
Cellulophaga sp. |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 15 | 65 | activity range, overview | Cellulophaga sp. |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
stable up to | Cellulophaga sp. |
| 40 | - |
purified recombinant His-tagged enzyme, pH 6.5, 1 h, 80% activity remaining | Cellulophaga sp. |
| 40 | - |
purified recombinant His-tagged enzyme, pH 6.5, 1 h, inactivation | Cellulophaga sp. |
| 50 | - |
purified recombinant His-tagged enzyme, pH 6.5, 1 h, 60% activity remaining | Cellulophaga sp. |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
- |
Cellulophaga sp. |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 11 | activity range, overview | Cellulophaga sp. |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | purified recombinant His-tagged enzyme, 45°C, 24 h, stable | Cellulophaga sp. |
| 6 | 9.6 | stable at | Cellulophaga sp. |
| 10 | - |
purified recombinant His-tagged enzyme, 45°C, 24 h, 60% activity remaining | Cellulophaga sp. |
| 11 | - |
purified recombinant His-tagged enzyme, 45°C, 24 h, inactivation | Cellulophaga sp. |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Cellulophaga sp. | sequence calculation | - |
4.79 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to glycohydrolase family 82, GH82, forming a deeply branched cluster in the phylogenetic tree, along with Celly_2571 and CgiA3, distinct from other iota-carrageenases | Cellulophaga sp. |
| evolution | the enzyme belongs to the glycosyl hydrolase family 82, GH82 | Cellulophaga sp. |
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