Tropane Alkaloid (TA) Producing Non-Plant Host Cells, and Methods of Making and Using the Same
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsChristina D. Smolke, Prashanth Srinivasan
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Abstract
Provided herein, among other things, is an engineered non-plant cell that produces a tropane alkaloid product, a precursor of a tropane alkaloid product, or a derivative of a tropane alkaloid product. A method for producing a tropane alkaloid, a precursor of a tropane alkaloid product, or a derivative of a tropane alkaloid product that makes use of the cell is also described.
Claims21
1-35. (canceled)
A method of producing a precursor of a tropane alkaloid product, a tropane alkaloid product, or a derivative of a tropane alkaloid product, the method comprising:
- (a) providing an engineered non-plant cell that produces the precursor of a tropane alkaloid product, the tropane alkaloid product, or the derivative of a tropane alkaloid product, wherein the engineered non-plant cell comprises:
- (i) a heterologous coding sequence encoding hyoscyamine dehydrogenase (HDH) within a pathway for producing the precursor, product, or derivative;
- and
- (ii) one or more alterations to one or more endogenous metabolic pathways or regulatory mechanisms selected from the group consisting of endogenous arginine metabolism, endogenous phenylalanine and phenylpropanoid metabolism, endogenous polyamine regulatory mechanisms and metabolism, endogenous acetate metabolism, and endogenous glycoside metabolism;
- (b) culturing the engineered non-plant cell in a medium comprising nutrients and water under conditions sufficient to produce the precursor, product, or derivative;
- and
- (c) recovering the precursor, product, or derivative from the engineered non-plant cell or the medium.
A method for forming a product stream comprising a tropane alkaloid product, a precursor of a tropane alkaloid product, or a derivative of a tropane alkaloid product, the method comprising:
- (a) providing to a reactor:
- (i) engineered non-plant cells comprising a heterologous coding sequence encoding hyoscyamine dehydrogenase (HDH) within a biosynthetic pathway for producing the tropane alkaloid product, the precursor, or the derivative, and
- (ii) a feedstock comprising nutrients and water;
- (b) fermenting the engineered non-plant cells in the reactor under conditions sufficient to produce a culture comprising the tropane alkaloid product, the precursor, or the derivative and cellular material;
- and
- (c) separating the tropane alkaloid product, the precursor, or the derivative from the cellular material to produce the product stream comprising the tropane alkaloid product, the precursor, or the derivative,
- wherein the engineered non-plant cells comprise one or more alterations to one or more endogenous metabolic pathways or regulatory mechanisms selected from the group consisting of endogenous arginine metabolism, endogenous phenylalanine and phenylpropanoid metabolism, endogenous polyamine regulatory mechanisms and metabolism, endogenous acetate metabolism, and endogenous glycoside metabolism.
Dependencies are inferred from the claim text.