Cloned (Comment) | Organism |
---|---|
functional coexpression of pinoresinol-lariciresinol reductase from Forsythia intermedia (FiPLR), secoisolariciresinol dehydrogenase from Podophyllum pleianthum (PpSDH), and CYP719A23 together with a suitable NADPH-dependent reductase to ensure P450 activity in Escherichia coli. Escherichia coli is cotransformed with two plasmids, generating a modular coexpression system: a pCDFDuet-1 vector harboring the genes encoding for FiPLR and PpSDH (FiPLR-PpSDH module) and either a pETDuet-1 or pET28a-(+) vector harboring P450-redox partner combinations (P450-module), method, overview | Dysosma pleiantha |
Protein Variants | Comment | Organism |
---|---|---|
additional information | assembly of plant enzymes in Escherichia coli for the production of the valuable (-)-podophyllotoxin precursor (-)-pluviatolide. (-)-Pluviatolide is considered a crossroad compound in lignan biosynthesis, because the methylenedioxy bridge in its structure, resulting from the oxidation of (-)-matairesinol, channels the biosynthetic pathway toward the microtubule depolymerizer (-)-podophyllotoxin. This oxidation reaction is catalyzed with high regio- and enantioselectivity by a cytochrome P450 monooxygenase from Sinopodophyllum hexandrum (CYP719A23), which is expressed and optimized regarding redox partners in Escherichia coli. Pinoresinol-lariciresinol reductase from Forsythia intermedia (FiPLR), secoisolariciresinol dehydrogenase from Podophyllum pleianthum (PpSDH), and CYP719A23 are coexpressed together with a suitable NADPH-dependent reductase to ensure P450 activity, allowing for four sequential biotransformations without intermediate isolation. By using an Escherichia coli strain coexpressing the enzymes originating from four plants, (+)-pinoresinol is efficiently converted, allowing the isolation of enantiopure (-)-pluviatolide at a concentration of 137 mg/l (enantiomeric excess over 99% with 76% isolated yield), reaction scheme and method, overview | Dysosma pleiantha |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(-)-secoisolariciresinol + 2 NAD+ | Dysosma pleiantha | - |
(-)-matairesinol + 2 NADH + 2 H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dysosma pleiantha | A0A0B4KYE1 | Dysosma pleiantha | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(-)-secoisolariciresinol + 2 NAD+ | - |
Dysosma pleiantha | (-)-matairesinol + 2 NADH + 2 H+ | - |
? | |
(-)-secoisolariciresinol + 2 NAD+ | the enantiomeric purity of formed (-)-matairesinol is over 99.9% in the reaction with the recombinant enzyme when tested with racemic secoisolariciresinol, high enantioselectivity, LC/MS and chiral HPLC analysis | Dysosma pleiantha | (-)-matairesinol + 2 NADH + 2 H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PpSDH | - |
Dysosma pleiantha |
SSDH | - |
Dysosma pleiantha |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Dysosma pleiantha |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Dysosma pleiantha |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Dysosma pleiantha |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is involved in the enzymatic cascade for (-)-podophyllotoxin biosynthesis | Dysosma pleiantha |