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Literature summary for 4.2.3.B68 extracted from
- Two diterpene synthases for spiroalbatene and cembrene A from allokutzneria albata (2018), Angew. Chem. Int. Ed. Engl., 57, 3238-3241 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| phylogenetic tree, recombinant expression of wild-type and mutant enzymes in Escherichia coli | Allokutzneria albata |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E179K | site-directed mutagenesis, inactive mutant | Allokutzneria albata |
| E179Q | site-directed mutagenesis, the mutation results in strongly reduced activity with both metal cations compared to wild-type | Allokutzneria albata |
| E193D | site-directed mutagenesis, the mutation results in reduced activity with Mg2+ compared to wild-type, or a complete loss of activity with Mn2+ | Allokutzneria albata |
| L72A | site-directed mutagenesis, the mutant variant of enzyme SaS gives no soluble protein | Allokutzneria albata |
| P201T | site-directed mutagenesis, the mutant variant of SaS shows significantly reduced activity with both metal cofactors compared to wild-type | Allokutzneria albata |
| P65A | site-directed mutagenesis, the mutation yields wild-type levels of enzyme, but the activity drops significantly with Mg2+ and Mn2+ | Allokutzneria albata |
| Q160E | site-directed mutagenesis, the mutant enzyme is well expressed, but shows no activity with either Mg2+ or Mn2+ | Allokutzneria albata |
| R145K | site-directed mutagenesis, the mutation results in reduced activity with Mg2+ compared to wild-type, or a complete loss of activity with Mn2+ | Allokutzneria albata |
| R145M | site-directed mutagenesis, the mutant variant shows only a slight decrease in activity for both metal cofactors compared to wild-type | Allokutzneria albata |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | activates | Allokutzneria albata |  |
| Mn2+ | activates | Allokutzneria albata | |
| additional information | influence of the length of this salt bridge, likely by alteration of the tilt angle between helices G and F on the active-site architecture, with consequences for metal binding. A short salt bridge with participation of Asp seems to favor catalysis with Mn2+ while a long salt bridge with Glu fosters Mg2+ binding | Allokutzneria albata |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| geranylgeranyl diphosphate | Allokutzneria albata | - |
spiroalbatene + diphosphate | - |
? | |
| geranylgeranyl diphosphate | Allokutzneria albata DSM 44149 | - |
spiroalbatene + diphosphate | - |
? | |
| additional information | Allokutzneria albata | for spiroalbatene synthase, the pH- and Mn2+-dependent formation of the side product thunbergol is observed, which is biosynthetically linked to spiroalbatene | ? | - |
? | |
| additional information | Allokutzneria albata DSM 44149 | for spiroalbatene synthase, the pH- and Mn2+-dependent formation of the side product thunbergol is observed, which is biosynthetically linked to spiroalbatene | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Allokutzneria albata | - |
isolated from tropical soil | - |
| Allokutzneria albata DSM 44149 | - |
isolated from tropical soil | - |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| geranylgeranyl diphosphate = spiroalbatene + diphosphate | cyclization mechanism from GGPP to spiroalbatene involving 1,14-cyclization to cation, followed by a 1,2-hydride migration, another 1,2-hydride shift, and ring closure, followed by a third 1,2-hydride shift and cyclization, yielding a cyclopropane intermediate, and opening of the three-membered ring with concomitant ring closure | Allokutzneria albata |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | the enzyme seems to be not expressed in laboratory cultures | Allokutzneria albata | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| geranylgeranyl diphosphate | - |
Allokutzneria albata | spiroalbatene + diphosphate | - |
? | |
| geranylgeranyl diphosphate | - |
Allokutzneria albata DSM 44149 | spiroalbatene + diphosphate | - |
? | |
| additional information | for spiroalbatene synthase, the pH- and Mn2+-dependent formation of the side product thunbergol is observed, which is biosynthetically linked to spiroalbatene | Allokutzneria albata | ? | - |
? | |
| additional information | the defined stereochemical anchors at the deuterated carbon atoms are used to solve the absolute configuration of spiroalbatene by determining the relative configurations of the stereospecifically deuterated products. Enzyme SaS can potentially yield ent-(S)-(+)-cembrene A from intermediate A.. Product identification by NMR spectroscopic analysis. Development of a method that uses the stereoselectively deuterated precursors (S)- and (R)-(1-13C,1-2H)FPP and (S)- and (R)-(1-13C,1-2H)GPP to determine the absolute configurations of terpenes | Allokutzneria albata | ? | - |
? | |
| additional information | for spiroalbatene synthase, the pH- and Mn2+-dependent formation of the side product thunbergol is observed, which is biosynthetically linked to spiroalbatene | Allokutzneria albata DSM 44149 | ? | - |
? | |
| additional information | the defined stereochemical anchors at the deuterated carbon atoms are used to solve the absolute configuration of spiroalbatene by determining the relative configurations of the stereospecifically deuterated products. Enzyme SaS can potentially yield ent-(S)-(+)-cembrene A from intermediate A.. Product identification by NMR spectroscopic analysis. Development of a method that uses the stereoselectively deuterated precursors (S)- and (R)-(1-13C,1-2H)FPP and (S)- and (R)-(1-13C,1-2H)GPP to determine the absolute configurations of terpenes | Allokutzneria albata DSM 44149 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| SAS | - |
Allokutzneria albata |
| spiroalbatene synthase | - |
Allokutzneria albata |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | 8.2 | assay at | Allokutzneria albata |
General Information
| General Information | Comment | Organism |
|---|---|---|
| metabolism | spiroalbatene may be oxidized to an unknown product by a genetically clustered cytochrome P450 | Allokutzneria albata |
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