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US 11,072,613 B2Granted
Compositions and methods for making terpenoid indole alkaloids
AssigneeWillow Biosciences, Inc.
InventorsVincenzo De Luca, Yang Qu
Life of this patent
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Abstract
Methods that may be used for the manufacture of a class of chemical compounds known as terpenoid indole alkaloids, including tabersonine and catharanthine are provided. Compositions useful for the synthesis of terpenoid indole alkaloids, including tabersonine and catharanthine are also provided. The provided compounds are useful in the manufacture of chemotherapeutic agents.
Claims15
A method of making tabersonine, comprising:
- (a) providing a terpenoid indole alkaloid compound;
- and
- (b) contacting the terpenoid indole alkaloid compound with catalytic quantities of a set of enzymes selected from
- (A) reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- (B) geissoschizine oxidase (GO), reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- (C) geissoschizine synthase (GS), geissoschizine oxidase (GO), reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);or
- (D) strictosidine β-glucosidase (SGD), geissoschizine synthase (GS), geissoschizine oxidase (GO), reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- wherein,
- (i) strictosidine β-glucosidase (SGD) comprises SEQ ID NO:
- 1 or a polypeptide having a sequence that is at least 85% identical thereto;
- (ii) geissoschizine synthase (GS) comprises SEQ ID NO:
- 2, 37, 38, or 39, or a polypeptide having a sequence that is at least 85% identical thereto;
- (iii) geissoschizine oxidase (GO) comprises SEQ ID NO:
- 3, 40, 41, or 42, or a polypeptide having a sequence that is at least 85% identical thereto;
- (iv) reductase 1 (REDOX 1) comprises SEQ ID NO:
- 4,43, 44, or 45, or a polypeptide having a sequence that is at least 85% identical thereto;
- (v) reductase 2 (REDOX 2) comprises SEQ ID NO:
- 5, 46, 47, or 48, or a polypeptide having a sequence that is at least 85% identical thereto;
- (vi) stemmadenine acetyltransferase (SAT) comprises SEQ ID NO:
- 16, 49, 50 or 51, or a polypeptide having a sequence that is at least 85% identical thereto;
- and
- (vii) hydrolyase 2 (HL2) comprises SEQ ID NO:
- 7, 52, 53, 54, 55, or 56, or a polypeptide having a sequence that is at least 85% identical thereto;
- under reaction conditions permitting the catalysis of the terpenoid indole alkaloid compound to form tabersonine.
21 more elements
A method for preparing tabersonine, wherein the method comprises:
- (a) providing a chimeric nucleic acid sequence comprising as operably linked components:
- (i) one or more nucleic acid sequences encoding a set of polypeptides selected from
- (A) reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- (B) geissoschizine oxidase (GO), reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- (C) geissoschizine synthase (GS), geissoschizine oxidase (GO), reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- or
- (D) strictosidine β-glucosidase (SGD), geissoschizine synthase (GS), geissoschizine oxidase (GO), reductase 1 (REDOX 1), reductase 2 (REDOX 2), stemmadenine acetyltransferase (SAT), and hydrolase 2 (HL2);
- wherein,
- strictosidine β-glucosidase (SGD) comprises SEQ ID NO:
- 1 or a polypeptide having a sequence that is at least 85% identical thereto;
- geissoschizine synthase (GS) comprises SEQ ID NO:
- 2, 37, 38, or 39, or a polypeptide having a sequence that is at least 85% identical thereto;
- geissoschizine oxidase (GO) comprises SEQ ID NO:
- 3, 40, 41, or 42, or a polypeptide having a sequence that is at least 85% identical thereto;
- reductase 1 (REDOX 1) comprises SEQ ID NO:
- 4, 43, 44, or 45, or a polypeptide having a sequence that is at least 85% identical thereto;
- reductase 2 (REDOX 2) comprises SEQ ID NO:
- 5, 46, 47 or 48, or a polypeptide having a sequence that is at least 85% identical thereto;
- stemmadenine acetyltransferase (SAT) comprises SEQ ID NO:
- 16, 49, 50 or 51, or a polypeptide having a sequence that is at least 85% identical thereto;
- and
- hydrolyase 2 (HL2) comprises SEQ ID NO:
- 7, 52, 53, 54, 55, or 56, or a polypeptide having a sequence that is at least 85% identical thereto;
- and
- (ii) one or more nucleic acid sequences capable of controlling expression in a host cell;
- (b) introducing the chimeric nucleic acid sequence into a host cell and growing the host cell to produce the encoded polypeptides;
- and
- (c) recovering tabersonine.
25 more elements
Dependencies are inferred from the claim text.