Application | Comment | Organism |
---|---|---|
medicine | a Saccharomyces cerevisiae strain is engineered to express seven heterologous enzymes (Papaper somniferum norcoclaurine 6-O-methyltransferase (Ps6OMT), Papaver somniferum 3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase 2 (Ps4'OMT), Papapver somniferum coclaurine N-methyltransferase (PsCNMT), Papaver somniferum berberine bridge enzyme (PsBBE), Thalictrum flavum scoulerine 9-O-methyltransferase (TfS9OMT), Thalictrum flavum canadine synthase (TfCAS), and Arabidopsis thaliana cytochrome P450 reductase 1 (CPR)), resulting in protoberberine alkaloid production from a simple benzylisoquinoline alkaloid precursor. A number of strategies are implemented to improve flux through the pathway, including enzyme variant screening, genetic copy number variation, and culture optimization. This leads to an over 70-fold increase in canadine titer up to 1.8 mg/l. Increased canadine titers enable extension of the pathway to produce berberine, a major constituent of several traditional medicines in a microbial host. This strain is viable at pilot scale | Papaver somniferum |
synthesis | heterologous production of berberine and the optimization of the engineered biosynthetic pathway from rac-norlaudanosoline to (S)-canadine in yeast involving the recombinant berberine reductase | Papaver somniferum |
Cloned (Comment) | Organism |
---|---|
canadine-producing Saccharomyces cerevisiae strain harbors expression cassettes for seven heterologous enzymes: Papaper somniferum norcoclaurine 6-O-methyltransferase (Ps6OMT), Papaver somniferum 3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase 2 (Ps4'OMT), Papapver somniferum coclaurine N-methyltransferase (PsCNMT), Papaver somniferum berberine bridge enzyme (PsBBE), Thalictrum flavum scoulerine 9-O-methyltransferase (TfS9OMT), Thalictrum flavum canadine synthase (TfCAS), and Arabidopsis thaliana cytochrome P450 reductase 1 (CPR). The expression cassettes for the methyltransferases Ps6OMT, PsCNMT, and Ps4'OMT and the cytochrome P450 reductase CPR were chromosomally integrated, TfS9OMT and TfCAS are expressed from a high-copy plasmid, and PsBBE is expressed from a second high-copy plasmid | Papaver somniferum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | recombinant coexpression of seven enzymes, including the three membrane-bound enzymes: the flavin-dependent oxidase berberine bridge enzyme, the cytochrome P450 canadine synthase, and a cytochrome P450 reductase, in Saccharomyces cerevisiae results in protoberberine alkaloid production from a simple benzylisoquinoline alkaloid precursor rac-norlaudanosoline. Improvement of flux through the pathway, by methods including enzyme variant screening, genetic copy number variation, and culture optimization, leading to an over 70fold increase in canadine titer up to 1.8 mg/l | Papaver somniferum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | membrane-bound | Papaver somniferum | 16020 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-reticuline + O2 | Papaver somniferum | - |
(S)-scoulerine + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Papaver somniferum | P93479 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-reticuline + O2 | - |
Papaver somniferum | (S)-scoulerine + H2O2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BBE | - |
Papaver somniferum |
BBE1 | - |
Papaver somniferum |
berberine bridge enzyme | - |
Papaver somniferum |
flavin-dependent oxidase | - |
Papaver somniferum |
reticuline oxidase | - |
Papaver somniferum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Papaver somniferum |