Compositions and methods for making (R)-reticuline and precursors thereof
Methods that may be used for the manufacture of the chemical compound (R)-Reticuline and synthesis precursors thereof. Compositions useful for the synthesis (R)-Reticuline and synthesis precursors are also provided.
1. A method for preparing (R)-Reticuline or an (R)-Reticuline precursor, the method comprising:
(a) introducing into a non-plant cell a chimeric nucleic acid sequence that encodes a polypeptide haying an amino acid sequence that corresponds to at least 85% of the sequence identity to the polypeptide encoded by SEQ ID NO: 322, comprising as operably linked components:
(i) a first nucleic acid sequence encoding CYP450;
(ii) a second nucleic acid sequence encoding AKR; and
(iii) one or more nucleic acid sequence capable of controlling expression in a host cell;
(b) growing the host cell to produce CYP450, AKR, and (R)-Reticuline or the (R)-Reticuline precursor.
2. The method of claim 1 , wherein the non-plant cell is a yeast cell.
3. The method of claim 1 , wherein the non-plant cell is a bacteria cell.
4. The method of claim 1 , wherein the chimeric nucleic acid sequence that encodes a polypeptide having an amino acid sequence that corresponds to at least 90% of the sequence identity to the polypeptide encoded by SEQ ID NO: 322.
5. The method of claim 1 , wherein the chimeric nucleic acid sequence that encodes a polypeptide having an amino acid sequence that corresponds to at least 95% of the sequence identity to the polypeptide encoded by SEQ ID NO: 322.
6. The method of claim 1 , wherein the host cell produces (S)-Reticuline.
7. The method of claim 1 , wherein the host cell produces (R)-Reticuline.
8. The method of claim 1 , wherein the host cell produces at least one compound selected from the group consisting of (S)-Coclaurine, (S)-N-Methylcoclaurine, (S)-3′-Hydroxy-N-methylcoclaurine, (R)-N-Methylcoclaurine, (R)-3′-Hydroxy-N-methylcoclaurine, and (S)-Reticuline.
9. The method of claim 1 , wherein the host cell produces at least one (R)-Reticuline precursor selected from the group consisting of (R)-N-Methylcoclaurine or (R)-3′-Hydroxy-N-methylcoclaurine.
10. The method of claim 1 , wherein growing the host cell comprises:
culturing the host cell in a cellular growth medium, wherein the cellular growth medium comprises at least one of (S)-Norcoclaurine, (S)--N-Methylnorcoclaurine, (S)-Norlaudanosaline, (S)--N-Methylnorlaudanosaline, (S)-Coclaurine, (S)-N-Methylcoclaurine, (S)-3′-Hydroxycoclaurine, (S)-3′-Hydroxy-N-methylcoclaurine, (S)-Higenamine, (S)--N -Methylhigenamine, (S)-Laudanosoline, (S)-Norreticuline, and (S)-Orientaline.