Compositions and methods for producing benzylisoquinoline alkaloids
AssigneeCalifornia Institute of Technology
InventorsChristina D. SMOLKE, Kristy M. HAWKINS
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Abstract
The present invention relates to host cells that produce compounds that are characterized as benzylisoquinolines, as well as select precursors and intermediates thereof. The host cells comprise one, two or more heterologous coding sequences wherein each of the heterologous coding sequences encodes an enzyme involved in the metabolic pathway of a benzylisoquinoline, or its precursors or intermediates from a starting compound. The invention also relates to methods of producing the benzylisoquinoline, as well as select precursors and intermediates thereof by culturing the host cells under culture conditions that promote expression of the enzymes that produce the benzylisoquinoline or precursors or intermediates thereof.
Claims11
-37. (canceled)
A method of producing an alkaloid compound in a host cell, the method comprising:
- (a) providing a first chimeric nucleic acid sequence comprising as operably linked components:
- (i) a first polynucleotide obtainable from a first plant capable of producing an alkaloid compound encoding a first enzyme capable of catalyzing a chemical reaction that converts a first alkaloid compound into a second alkaloid compound;
- and
- (ii) one or more polynucleotides capable of controlling expression in a cell;
- (b) providing a second chimeric nucleic acid sequence comprising as operably linked components:
- (i) a second polynucleotide obtainable from a second plant capable of producing an alkaloid compound encoding a second enzyme capable of catalyzing a chemical reaction that converts the second alkaloid compound into a third alkaloid compound;
- and
- (ii) one or more polynucleotides capable of controlling expression in a cell;
- (c) introducing the first and second chimeric nucleic acid sequence into a cell;
- and
- (d) growing the cell to produce the first and second enzyme and the second and third alkaloid compound;
- and
- wherein the first and second polynucleotide are non-homologous and wherein the third alkaloid compound is produced in the cell at a level that is in excess of the level of alkaloid compound produced when a homologous first and second polynucleotide are used.
A method of producing an alkaloid compound in an engineered non-plant cell, the method comprising:
- (a) providing a first heterologous coding sequence comprising as operably linked components:
- (i) a first polynucleotide obtainable from a first plant capable of producing an alkaloid compound encoding a first enzyme capable of catalyzing a chemical reaction that converts a first alkaloid compound into a second alkaloid compound;
- and
- (ii) one or more polynucleotides capable of controlling expression in the engineered non-plant cell;
- (b) providing a second heterologous coding sequence comprising as operably linked components:
- (i) a second polynucleotide obtainable from a second plant capable of producing an alkaloid compound encoding a second enzyme capable of catalyzing a chemical reaction that converts the second alkaloid compound into a third alkaloid compound;
- and
- (ii) one or more polynucleotides capable of controlling expression in the engineered non-plant cell;
- (c) introducing the first and second heterologous coding sequences into the engineered non-plant cell;
- and
- (d) growing the cell to produce the first and second enzymes and the second and third alkaloid compounds;
- and
- wherein the first and second polynucleotides are non-homologous and wherein the third alkaloid compound is produced in the engineered non-plant cell at a level that is in excess of the level of alkaloid compound produced when a homologous first and second polynucleotide are used.
Dependencies are inferred from the claim text.