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Literature summary for 4.2.1.178 extracted from
- Structural and functional basis of difructose anhydride III hydrolase, which sequentially converts inulin using the same catalytic residue (2018), ACS Catal., 8, 10683-10697 .
No PubMed abstract available
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Pseudarthrobacter chlorophenolicus |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| hanging drop vapor diffusion method, using 0.1 M sodium malonate (pH 4.2) and 10% (w/v) PEG 3350 | Pseudarthrobacter chlorophenolicus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C387A | the mutant shows about 1.85fold increased activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
| C387A/E210A | inactive | Pseudarthrobacter chlorophenolicus |
| D177A | inactive | Pseudarthrobacter chlorophenolicus |
| D177N | inactive | Pseudarthrobacter chlorophenolicus |
| D199A | inactive | Pseudarthrobacter chlorophenolicus |
| D199N | inactive | Pseudarthrobacter chlorophenolicus |
| E210A | inactive | Pseudarthrobacter chlorophenolicus |
| E210Q | inactive | Pseudarthrobacter chlorophenolicus |
| E307A | inactive | Pseudarthrobacter chlorophenolicus |
| F207A | the mutant shows 67.72% activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
| F256A | the mutant shows 17.65% activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
| F80A | the mutant shows 8.61% activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
| I85A | the mutant shows 14.75% activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
| Q222A | inactive | Pseudarthrobacter chlorophenolicus |
| Q391A | inactive | Pseudarthrobacter chlorophenolicus |
| R134A | inactive | Pseudarthrobacter chlorophenolicus |
| R258A | inactive | Pseudarthrobacter chlorophenolicus |
| W309F | the mutant shows 10.32% activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
| Y163A | the mutant shows 8.09% activity with D-fructofuranose 2',1:2,3'-dianhydride compared to the wild type enzyme | Pseudarthrobacter chlorophenolicus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 98.06 | - |
alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride | mutant enzyme C387A, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |  |
| 121.3 | - |
alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride | wild type enzyme, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 139000 | - |
gel filtration | Pseudarthrobacter chlorophenolicus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride + H2O | Pseudarthrobacter chlorophenolicus | - |
inulobiose | - |
r | |
| inulobiose | Pseudarthrobacter chlorophenolicus | - |
alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride + H2O | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudarthrobacter chlorophenolicus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| HiTrap Q Sepharose column chromatography and Superdex 200 gel filtration | Pseudarthrobacter chlorophenolicus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 5.17 | - |
mutant enzyme W309F, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 8.19 | - |
mutant enzyme Y163A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 8.71 | - |
mutant enzyme F80A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 10.45 | - |
mutant enzyme S84A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 14.93 | - |
mutant enzyme I85A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 17.87 | - |
mutant enzyme F256A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 67.72 | - |
mutant enzyme F207A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 101.25 | - |
wild type enzyme with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
| 187.51 | - |
mutant enzyme C387A, with alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride as substrate, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride + H2O | - |
Pseudarthrobacter chlorophenolicus | inulobiose | - |
r | |
| inulobiose | - |
Pseudarthrobacter chlorophenolicus | alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride + H2O | - |
r | |
| additional information | the enzyme is also capable of converting inulin to alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride (inulin fructotransferase activity) | Pseudarthrobacter chlorophenolicus | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotrimer | 3 * 47000, SDS-PAGE | Pseudarthrobacter chlorophenolicus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DFA-IIIase | - |
Pseudarthrobacter chlorophenolicus |
| difructose anhydride III hydrolase | - |
Pseudarthrobacter chlorophenolicus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
- |
Pseudarthrobacter chlorophenolicus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 80 | about 40% activity at 30°C, about 60% activity at 40°C, about 95% activity at 50°C, 100% activity at 55°C, about 95% activity at 60°C, about 50% activity at 65°C, about 40% activity at 70°C, about 30% activity at 80°C | Pseudarthrobacter chlorophenolicus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
- |
Pseudarthrobacter chlorophenolicus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 7 | more than 70% activity between pH 5.5 and 7.0 | Pseudarthrobacter chlorophenolicus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 4.61 | - |
alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride | wild type enzyme, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus | |
| 8.59 | - |
alpha-D-fructofuranose-beta-D-fructofuranose 2',1:2,3'-dianhydride | mutant enzyme C387A, at pH 6.5 and 55°C | Pseudarthrobacter chlorophenolicus | |
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