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US 2026/0286406 A1Published application
Paclitaxel (taxol) biosynthesis pathway
AssigneesMax-Planck-Gesellschaft zur Forderung der Wissenschaften e.V., Center of Plant Systems Biology and Biotechnology
InventorsAlisdair R. Fernie, Youjun Zhang
Life of this patent
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Abstract
The present invention relates to the Paclitaxel (taxol) biosynthesis pathway and in particular to methods for producing 4β,20-taxadiene, 10-deacetylbaccatin III, Baccatin III or β-phenylalanoyl-coA as well as corresponding uses. The present invention further relates to a vector, recombinant organism, a tissue, a cell, or an organelle comprising the enzymes being required for the production of 4β,20-taxadiene, 10-deacetylbaccatin III, Baccatin III or β-phenylalanoyl-coA.
Claims15
A method for producing 4β,20-taxadiene comprising
- (a) enzymatically converting taxa-4(5),11(12)-diene with a C4β,C20-epoxidase selected from
- (i) a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 3;
- (ii) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 4;
- (iii) a polypeptide having an amino acid sequence being at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 3;
- (iv) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:
- 4;
- and
- (v) a fragment of the polypeptide of any of (i) to (iv) comprising at least 200 amino acids
- thereby producing 4β,20-taxadiene.
9 more elements
A method for producing 10-deacetylbaccatin III comprising
- (a) enzymatically converting 9-di-10-deacetylbaccatin III with an T9α-oxidase selected from
- (i) a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 25, 39, 41 or 43;
- (ii) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 26, 40, 42 or 44;
- (iii) a polypeptide having an amino acid sequence being at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 25, 39, 41 or 43;
- (iv) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:
- 26, 40, 42 or 44;
- and
- (v) a fragment of the polypeptide of any of (i) to (iv) comprising at least 300 amino acids
- thereby producing 10-deacetylbaccatin III.
9 more elements
A method for producing β-phenylalanoyl-coA (i) from β-phenylalanine by β-phenylalanine coenzyme A ligase (PLC, EC 6.2.1.30), or (ii) from L-phenylalanine by phenylalanine aminomutase (PAM, EC 5.4.3.11) and β-phenylalanine coenzyme A ligase (EC 6.2.1.30),
- wherein the β-phenylalanine coenzyme A ligase is selected from
- (i) a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 33;
- (ii) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 34;
- (iii) a polypeptide having an amino acid sequence being at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 33;
- (iv) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:
- 34;
- and
- (v) a fragment of the polypeptide of any of (i) to (iv) comprising at least 300 amino acids;
- and
- preferably wherein the phenylalanine aminomutase is selected from
- (i) a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 31;
- (ii) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 32;
- (iii) a polypeptide having an amino acid sequence being at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 31;
- (iv) a polypeptide being encoded by a nucleic acid molecule comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:
- 32;
- and
- (v) a fragment of the polypeptide of any of (i) to (iv) comprising at least 500 amino acids.
20 more elements
A vector comprising
- (I)
- a nucleic acid molecule encoding a C4β,C20-epoxidase, which nucleic acid molecule is
- (a) a nucleic acid molecule encoding a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 3;
- (b) a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 4;
- (c) a nucleic acid molecule comprising or consisting of a nucleic acid molecule encoding a polypeptide having the activity of a catalase the amino acid sequence of which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 3;
- (d) a nucleic acid molecule encoding a polypeptide comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:
- 4;
- (e) a fragment of the nucleic acid molecule of any of (i) to (iv) comprising at least 600 nucleic acids;
- or
- (f) the nucleic acid sequence of any of (a) to (e) wherein T is U;
- and/or
- (II)
- a nucleic acid molecule encoding a T9α-oxidase, which nucleic acid molecule is
- (a) a nucleic acid molecule encoding a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 25, 39, 41 or 43;
- (b) a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 26, 40, 42 or 44;
- (c) a nucleic acid molecule comprising or consisting of a nucleic acid molecule encoding a polypeptide having the activity of a catalase the amino acid sequence of which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 25, 39, 41 or 43;
- (d) a nucleic acid molecule encoding a polypeptide comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:
- 26, 40, 42 or 44;
- (e) a fragment of the nucleic acid molecule of any of (i) to (iv) comprising at least 900 nucleic acids, or
- (f) the nucleic acid sequence of any of (a) to (e) wherein T is U;
- and/or
- (III)
- a nucleic acid molecule encoding a β-phenylalanine coenzyme A ligase (PLC, EC 6.2.1.30), which nucleic acid molecule is
- (a) a nucleic acid molecule encoding a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO:
- 33;
- (b) a nucleic acid molecule comprising or consisting of the nucleotide sequence of SEQ ID NO:
- 34;
- (c) a nucleic acid molecule comprising or consisting of a nucleic acid molecule encoding a polypeptide having the activity of a catalase the amino acid sequence of which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the amino acid sequence of SEQ ID NO:
- 33;
- (d) a nucleic acid molecule encoding a polypeptide comprising or consisting of a nucleotide sequence which is at least 80% identical, preferably at least 90% and most preferably at least 95% identical to the nucleotide sequence of SEQ ID NO:34;
- (e) a fragment of the nucleic acid molecule of any of (i) to (iv) comprising at least 900 nucleic acids, or
- (f) the nucleic acid sequence of any of (a) to (e) wherein T is U.
35 more elements
Dependencies are inferred from the claim text.