Cells and methods for producing rhamnolipids

1. A genetically modified Pseudomonas putida cell, which is able to form at least one rhamnolipid of general formula (I),

wherein

m=2, 1 or 0,

n=1 or 0,

R 1 and R 2 are organic residues having 2 to 24 carbon atoms,

wherein said cell has been genetically modified such that, compared to its wild-type, the cell has an increased activity of at least one of the enzymes E 1 , E 2 and E 3 , wherein the enzyme E 1 has at least 95% amino acid identity to SEQ ID NO: 2, the enzyme E 2 has at least 95% amino acid identity to SEQ ID NO: 4, and the enzyme E 3 has at least 95% amino acid identity to SEQ ID NO: 6;

and wherein said cell is also genetically modified such that, compared to its wild-type, the cell has increased activity of at least one enzyme selected from the group consisting of: E 4 , which has at least 95% amino acid identity to SEQ ID NO: 10, E 5 , which has at least 95% amino acid identity to SEQ ID NO: 12, E 6 , which has at least 95% amino acid identity to SEQ ID NO: 16, and E 7 , which has at least 95% amino acid identity to SEQ ID NO: 14.

2. The genetically modified cell of claim 1 , wherein said cell has increased activities of an enzyme combination selected from

E 1 E 2 , E 2 E 3 and E 1 E 2 E 3 .

3. The genetically modified cell of claim 1 , wherein said cell has an increased activity of the enzyme combination

E 1 E 2 E 3 and

n is =1.

4. The genetically modified cell of claim 1 , wherein the wild-type of said cell forms polyhydroxyalkanoates having chain lengths of C 6 to C 16 .

5. The genetically modified cell of claim 4 , wherein said cell, compared to its wild-type, has a decreased activity of at least one enzyme E 9 or E 10 ,

wherein E 9 has an amino acid sequence with at least 95% identity to the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 32, and

E 10 has an amino acid sequence with at least 95% identity to the amino acid sequence of SEQ ID NO: 34 or SEQ ID NO: 36.

6. The genetically modified cell of claim 1 , wherein said cell is selected from the group consisting of P. putida GPp121, P. putida GPp122, P. putida GPp123, P. putida GPp124 and P. putida GPp104, P. putida KT42C1, P. putida KTOY01 and P. putida KTOY02.

7. The genetically modified cell of claim 1 , wherein said cell, compared to its wild-type, has increased activity of each of the enzymes

E 4 ,

E 5 ,

E 6 , and

E 7 .

8. The genetically modified cell of claim 1 , wherein said cell, compared to its wild-type, further has an increased activity of an enzyme E 8 , which catalyzes rhamnolipid export from the cell into the surrounding medium.

9. The genetically modified cell of claim 1 , wherein said genetic modification comprises introduction into said cell of at least one vector comprising a sequence selected from:

a sequence with at least 95% identity to SEQ ID NO: 1,

a sequence with at least 95% identity to SEQ ID NO: 3, and

a sequence with at least 95% identity to SEQ ID NO: 5.

10. The genetically modified cell of claim 9 , wherein the vector comprises the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 47.

11. The genetically modified cell of claim 9 , wherein said at least one vector comprises:

a sequence with at least 95% identity to SEQ ID NO: 1, a sequence with at least 95% identity to SEQ ID NO: 3, and a sequence with at least 95% identity to SEQ ID NO: 5.

12. The genetically modified cell of claim 9 , wherein said at least one vector further comprises a sequence selected from:

a sequence with at least 95% identity to SEQ ID NO: 9, a sequence with at least 95% identity to SEQ ID NO: 11, a sequence with at least 95% identity to SEQ ID NO: 13, and a sequence with at least 95% identity to SEQ ID NO: 15.

13. A method for producing rhamnolipids of general formula (I)

wherein

m=2, 1 or 0,

n=1 or 0

R 1 and R 2 are organic residues having 2 to 24 carbon atoms, wherein said method comprises

I) contacting a genetically modified Pseudomonas putida cell according to claim 1 with a medium containing a carbon source; and

II) culturing the cell under conditions in which the cell forms rhamnolipids from the carbon source.

14. A vector comprising at least one nucleic acid sequence selected from the group consisting of SEQ ID NO: 41, and SEQ ID NO: 50.