EC Number   |
Protein Variants |
Reference |
|---|
   1.1.1.331 | K171A |
mutation in conserved catalytic triad, complete loss of activity |
720614 |
| 1.1.1.331 | more |
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 |
762561 |
| 1.1.1.331 | more |
podophyllotoxin biosynthesis pathway genes expression at low temperature. The low temperature enhances the podophyllotoxin accumulation in Dysosma versipellis |
763661 |
| 1.1.1.331 | more |
the two genes, termed plr-PpH and sdh-PpH, encoding pinoresinol-lariciresinol reductase (PLR) and secoisolariciresinol dehydrogenase (SDH), are linked to form two bifunctional fusion genes, plr-sdh and sdh-plr, which are expressed in Escherichia coli and purified. The proteins are linked via a (GGGGS)4 protein linker to maintain flexibility. Bioconversion in vitro at 22°C for 60 min shows that the conversion efficiency of fusion protein PLR-SDH is higher than that of the mixture of recombinant PLR and reacombinant SDH. The percent conversion of (+)-pinoresinol to matairesinol is 49.8% using PLR-SDH and only 17.7% using a mixture of rPLR and rSDH. Conversion of (+)-pinoresinol by fusion protein SDH-PLR stops at the intermediate product, secoisolariciresinol. In vivo, (+)-pinoresinol is completely converted to matairesinol by living recombinant Escherichia coli expressing PLR-SDH without addition of cofactors |
741681 |
| 1.1.1.331 | S153A |
mutation in conserved catalytic triad, severe reduction of activity |
720614 |
| 1.1.1.331 | Y167A |
mutation in conserved catalytic triad, complete loss of activity |
720614 |
