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Literature summary for 3.2.1.157 extracted from
- Hyper-production and characterization of the iota-carrageenase useful for iota-carrageenan oligosaccharide production from a deep-sea bacterium, Microbulbifer thermotolerans JAMB-A94T, and insight into the unusual catalytic mechanism (2011), Mar. Biotechnol., 13, 411-422.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Bacillus subtilis | Microbulbifer thermotolerans |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D359N | the mutant shows reduced activity compared to the wild type enzyme. Asp359 is not critical for catalysis | Microbulbifer thermotolerans |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Cs+ | in the presence of 0.1 M CsCl, 63% of maximal activity is observed | Microbulbifer thermotolerans |  |
| K+ | in the presence of 0.1 M KCl, 86% of maximal activity is observed | Microbulbifer thermotolerans | |
| Li+ | in the presence of 0.1 M LiCl, 42% of maximal activity is observed | Microbulbifer thermotolerans | |
| Na+ | the enzyme requires the Na+ ion for full activity, maximal activity is observed in the presence of 0.1 M NaCl. In buffer containing 10 mM NaCl, the enzyme activity decreases to 28% during 16 h incubation on ice | Microbulbifer thermotolerans | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 54000 | - |
gel filtration | Microbulbifer thermotolerans |
| 55000 | - |
1 * 55000, SDS-PAGE | Microbulbifer thermotolerans |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Microbulbifer thermotolerans | E3W9G3 | - |
- |
| Microbulbifer thermotolerans JAMB-A94T | E3W9G3 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| ammonium sulfate precipitation, butyl-Toyopearl column chromatography, heparin affinity column chromatography, and Superdex 200 gel filtration | Microbulbifer thermotolerans |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 7.8 | - |
culture supernatant, at pH 7.5 and 50°C | Microbulbifer thermotolerans |
| 80.6 | - |
after 10.3fold purification, at pH 7.5 and 50°C | Microbulbifer thermotolerans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| iota-carrageenan + H2O | the enzyme cleaves beta-1,4 linkages in iota-carrageenan to produce a high ratio of iota-carrageenan tetramer, more than 75% of the total product | Microbulbifer thermotolerans | iota-neocarratetraose sulfate + iota-neocarrahexaose sulfate | - |
? | |
| iota-carrageenan + H2O | the enzyme cleaves beta-1,4 linkages in iota-carrageenan to produce a high ratio of iota-carrageenan tetramer, more than 75% of the total product | Microbulbifer thermotolerans JAMB-A94T | iota-neocarratetraose sulfate + iota-neocarrahexaose sulfate | - |
? | |
| additional information | the enzyme does not cleave the beta-1,4 linkages in kappa- or lambda-carrageenan | Microbulbifer thermotolerans | ? | - |
? | |
| additional information | the enzyme does not cleave the beta-1,4 linkages in kappa- or lambda-carrageenan | Microbulbifer thermotolerans JAMB-A94T | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 55000, SDS-PAGE | Microbulbifer thermotolerans |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CgiA | - |
Microbulbifer thermotolerans |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
in 50 mM Tris-HCl, pH 7.5, containing 0.1 M NaCl and 1 mM CaCl2 | Microbulbifer thermotolerans |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | 60 | - |
Microbulbifer thermotolerans |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | - |
the enzyme is stable up to 45°C in 50 mM Tris/HCl, pH 7.5, containing 0.1 M NaCl and 1 mM CaCl2 for 1 h. Incubation at 45°C inactivates the enzyme completely within 5 min at either condition of low NaCl concentration (less than 10 mM) or presence of 10 mM EDTA | Microbulbifer thermotolerans |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Microbulbifer thermotolerans |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 10 | in a range of pH 7.0-10.0, the enzyme retains more than 80% of the original activity | Microbulbifer thermotolerans |
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