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1. (WO2019064243) HETEROLOGOUS BIOSYNTHESIS OF NODULISPORIC ACID
注意: このテキストは、OCR 処理によってテキスト化されたものです。法的な用途には PDF 版をご利用ください。

What we claim is:

1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of NodW (SEQ ID NO:3), NodR (SEQ ID NO:6), NodJ (SEQ ID NO:21), NodYl (SEQ ID NO:27), NodY2 (SEQ ID NO:36), NodX (SEQ ID NO:9), NodM (SEQ ID NO: 12), NodB (SEQ ID NO: 15), NodO (SEQ ID NO: 18), NodC (SEQ ID NO: 24), NodD2 (SEQ ID NO: 30), NodDl (SEQ ID NO:33), NodZ (SEQ ID NO: 39), and NodS (SEQ ID NO: 50) or a functional variant or fragment thereof.

2. An isolated polynucleotide encoding a polypeptide of claim 1.

3. An isolated polynucleotide comprising at least 70% preferably at least 80%, preferably at least 90%, preferably at least 95%, preferably 99% nucleic acid sequence identity to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1, 2,

4. 5, 19, 20, 25, 26, 34, 35, 7, 8, 10, 11, 13, 14, 16, 17, 22, 23, 28, 29, 31, 32, 37, 38, 48 and 49.

4. A transcription unit (TU) comprising at least one isolated polynucleotide of claim 3.

5. A vector that encodes an isolated polypeptide of claim 1.

6. A vector comprising an isolated polynucleotide of claim 2 or claim 3, or a TU of cla im 4.

7. An isolated host cell comprising an isolated polypeptide of claim 1, an isolated polynucleotide of claim 2 or claim 3, a TU of claim 4 and/or a vector of claim 5 or claim 6.

8. A method of making at least one Nodulisporic Acid (NA) comprising heterologously expressing at least one isolated polypeptide of claim 1, isolated polynucleotide of claim 2 or claim 3, a TU of claim 4 and/or a vector of claim 5 or claim 6 in an isolated host cell.

9. An isolated polypeptide or functional fragment or variant thereof from Hypoxylon spp. that catalyzes a biochemical reaction in the biosynthetic pathway leading from 3-geranylgeranyl indole (GGI) 2 to NAA 10.

10. An isolated polynucleotide encoding at least one polypeptide or functional variant or fragment thereof from Hypoxylon spp. that catalyzes a biochemical reaction in the biosynthetic pathway leading from GGI 2 to NAA 10.

11. A method of making at least one Hypoxylon spp. polypeptide or functional variant or fragment thereof comprising heterologously expressing an isolated polynucleotide of cla im 2 or claim 3 or a vector of claim 5 or claim 6 in an isolated host cell.

12. A method of making at least one NA comprising heterologously expressing in an isolated host cell, at least one polypeptide that catalyzes a biochemical reaction in the biosynthetic pathway leading from GGI 2 to NAA 10.

13. An isolated host cell that expresses at least one heterologous polypeptide that catalyzes the transformation of a substrate in the biosynthetic pathway leading from GGI 2 to the formation of NAA 10.

14. An isolated host cell that produces by heterologous expression, at least one polypeptide involved in the biosynthetic pathway leading from GGI 2 to NAA 10.

15. A method of producing at least one NA comprising contacting a carbohydrate comprising substrate with a recombinant cell transformed with a nucleic acid that results in an increased level of activity of a polypeptide selected from the group consisting of SEQ ID NO: 3, 6, 21, 27, 36, 9, 12, 15, 18, 24, 30, 33, 39 and 50 or a functional variant or fragment thereof compared to the cell prior to transformation, such that the substrate is metabolized to at least one NA.

16. An isolated strain of Hypoxylon pulicicidum that comprises at least one

heterologous nucleic acid sequence encoding an enzyme in a biosynthetic pathway leading to NAA 10.

17. An isolated strain of Hypoxylon pulicicidum that expresses at least two different GGPPS enzymes.

18. An isolated strain of Hypoxylon pulicicidum that comprises a genetic modification that leads to an increased biosynthesis of NAA 10.

19. A method of making NAA 10 comprising expressing at least one heterologous nucleic acid sequence in Hypoxylon pulicicidum, wherein the at least one heterologous nucleic acid sequence encodes an enzyme in a biosynthetic pathway leading to NAA 10.