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US 10,190,141 B2Granted
Compositions and methods for making (R)-Reticuline and precursors thereof
AssigneeEpimeron Inc.
InventorsPeter James Facchini, Scott Cameron Farrow, Guillaume Arthur Welch Beaudoin
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Abstract
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.
Claims15
A method of making (R)-Reticuline or a precursor of (R)-Reticuline comprising
- (a) contacting a benzylisoquinoline derivative with a microorganism expressing a heterologous enzyme capable of converting the benzylisoquinoline derivative to (R)-Reticuline or an (R)-Reticuline precursor, wherein said enzyme is a fusion protein of CYP450 and AKR and comprises an amino acid sequence that is at least 85% identical to SEQ.ID NO:
- 323;
- and
- (b) growing said microorganism to produce (R)-Reticuline or a precursor of (R)-Reticuline.
1 more element
A method for preparing (R)-Reticuline or an (R)-Reticuline precursor comprising:
- (a) providing a chimeric nucleic acid sequence comprising as operably linked components:
- (i) a first nucleic acid sequence encoding a CYP450 polypeptide;
- (ii) a second nucleic acid sequence encoding an AKR polypeptide;
- and
- (iii) one or more nucleic acid sequences capable of controlling expression in a host cell;
- (b) introducing said chimeric nucleic acid sequence into said host cell and growing said host cell to produce CYP450 and AKR and to produce (R)-Reticuline or the (R)-Reticuline precursor, wherein said CYP450 and AKR form a fusion protein comprising an amino acid sequence that is at least 85% identical to SEQ.ID NO:
- 323;
- and
- (c) recovering (R)-Reticuline or the (R)-Reticuline precursor.
6 more elements
A recombinant expression vector suitable for expression in a host cell comprising as operably linked components:
- (i) a nucleic acid sequence capable of controlling expression in said host cell;
- and
- (ii) a nucleic acid sequence encoding CYP450 and AKR,
- wherein said CYP450 and AKR form a fusion protein comprising an amino acid sequence that is at least 85% identical to SEQ.ID NO:
- 323.
2 more elements
Dependencies are inferred from the claim text.