Host cells and methods for producing cinnamoyl anthranilate and analogs thereof
View Patent ↗The present invention provides for a method of producing a cinnamoyl anthranilate, or analog thereof, in a genetically modified host cell.
1. A culture medium comprising: (A) a genetically modified host cell comprising (a) an hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT, EC 2.3.1.144) having an amino acid sequence at least 70% identical to SEQ ID NO:1 capable of catalyzing the formation of a cinnamoyl anthranilate, or analog thereof, from a cinnamoyl-CoA, or analog thereof, and an anthranilate, or analog thereof, and (b) a 4-coumarate:CoA ligase (4CL, EC 6.2.1.12) having an amino acid sequence at least 70% identical to SEQ ID NO:3 capable of catalyzing the formation of a cinnamic acid, or analog thereof, into a corresponding cinnamoyl-CoA thioester, or analog thereof; and (B) a compound having the following chemical structure:
wherein R 1 and R 2 are each H; R 3 is OCH 3 ; and R 4 is OCH 3 .
2. The culture medium of claim 1 , wherein the HCBT has the amino acid sequence of SEQ ID NO:1.
3. The culture medium of claim 1 , wherein the 4CL has the amino acid sequence of SEQ ID NO:3.
4. A method of producing a cinnamoyl anthranilate, or analog thereof, comprising: culturing the genetically modified host cell of claim 1 under a suitable condition such that the culturing results in the genetically modified host cell producing an cinnamoyl anthranilate, or analog thereof.
5. The method of claim 4 , further comprising isolating the cinnamoyl anthranilate, or analog thereof, from the host cell and/or culture medium.
6. The culture medium of claim 1 , wherein the HCBT has an amino acid sequence at least 80% identical to SEQ ID NO:1.
7. The culture medium of claim 6 , wherein the HCBT has an amino acid sequence at least 90% identical to SEQ ID NO:1.
8. The culture medium of claim 7 , wherein the HCBT has an amino acid sequence at least 95% identical SEQ ID NO:1.
9. The culture medium of claim 1 , wherein the 4CL has an amino acid sequence at least 80% identical to SEQ ID NO:3.
10. The culture medium of claim 9 , wherein the 4CL has an amino acid sequence at least 90% identical to SEQ ID NO:3.
11. The culture medium of claim 10 , wherein the 4CL has an amino acid sequence at least 95% identical SEQ ID NO:3.
12. The culture medium of claim 1 , wherein the host cell is a bacteria of a genus selected from the group consisting of Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsielia, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla, Synechococcus, Synechocystis , and Paracoccus.
13. The culture medium of claim 1 , wherein the host cell is a yeast.
14. The culture medium of claim 13 , wherein the yeast is Saccharomyces cerevisiae.