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Literature summary for 1.1.1.331 extracted from

  • Decembrino, D.; Ricklefs, E.; Wohlgemuth, S.; Girhard, M.; Schullehner, K.; Jach, G.; Urlacher, V.B.
    Assembly of plant enzymes in E. coli for the production of the valuable (-)-podophyllotoxin precursor (-)-pluviatolide (2020), ACS Synth. Biol., 9, 3091-3103 .
    View publication on PubMed
Search for more literature for 1.1.1.331

Cloned(Commentary)

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

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/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(-)-secoisolariciresinol + 2 NAD+ Dysosma pleiantha
-
 (-)-matairesinol + 2 NADH + 2 H+
-
 ?

Organism

Organism UniProt Comment Textmining
Dysosma pleiantha A0A0B4KYE1 Dysosma pleiantha
-

Substrates and Products (Substrate)

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

Synonyms Comment Organism
PpSDH
-
 Dysosma pleiantha
SSDH
-
 Dysosma pleiantha

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Dysosma pleiantha

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Dysosma pleiantha

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Dysosma pleiantha

General Information

General Information Comment Organism
metabolism the enzyme is involved in the enzymatic cascade for (-)-podophyllotoxin biosynthesis Dysosma pleiantha