Method for producing 4-aminocinnamic acid, and vector and host cell used in same

1. A method for producing 4-aminocinnamic acid from 4-nitrophenylalanine, comprising the steps of:

(1) converting 4-nitrophenylalanine to 4-nitrocinnamic acid by using a first enzyme which consists of an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence defined in SEQ ID NOs:1, 3, or 5 and which has the ability to convert 4-nitrophenylalanine to 4-nitrocinnamic acid; and

(2) converting 4-nitrocinnamic acid to 4-aminocinnamic acid by using a second enzyme which consists of an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence defined in SEQ ID NOs:7, 9, 11, 13, or 15 and which has the ability to convert 4-nitrocinnamic acid to 4-aminocinnamic acid.

2. The method according to claim 1 , wherein the conversion of step (1) is carried out with a first host cell which has been engineered to express the first enzyme.

3. The method according to claim 2 , wherein the first host cell is a microorganism cell.

4. The method according to claim 3 , wherein the microorganism is a bacterium.

5. The method according to claim 4 , wherein the conversion of step (1) is carried out via a resting-cell reaction using a resting bacterial cell as the first host cell.

6. The method according to claim 1 , wherein the conversion of step (2) is carried out by using a second host cell which expresses the second enzyme.

7. The method according to claim 6 , wherein the second host cell is a host cell engineered to express the second enzyme.

8. The method according to claim 6 , wherein the second host cell is a microorganism cell.

9. The method according to claim 8 , wherein the microorganism is a bacterium.

10. The method according to claim 9 , wherein the conversion of step (2) is carried out via a resting-cell reaction using a resting bacterial cell as the second host cell.

11. The method according to claim 5 , wherein the resting bacterial cell is selected from the group consisting of cultured cells, powdered cells, and immobilized cells.

12. The method according to claim 1 , wherein the conversion of step (2) is carried out at a pH of from 8 to 9.

13. A method for producing 4-aminocinnamic acid from glucose, comprising the steps of:

(a) producing phenylalanine from glucose;

(b) converting the phenylalanine obtained in step (a) to 4-nitrophenylalanine via nitration; and

(c) producing 4-aminocinnamic acid from the 4-nitrophenylalanine obtained in step (b) via a method according to claim 1 .

14. A method for producing 4-aminocinnamic acid from phenylalanine, comprising the steps of:

(b) converting phenylalanine to 4-nitrophenylalanine via nitration; and

(c) producing 4-aminocinnamic acid from the 4-nitrophenylalanine obtained in step (b) via a method according to claim 1 .

15. A method for producing 4-nitrocinnamic acid from 4-nitrophenylalanine, comprising:

using a first enzyme which consists of an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence defined in SEQ ID NOs:1, 3, or 5 and which has the ability to convert 4-nitrophenylalanine to 4-nitrocinnamic acid.

16. A method for producing 4-aminocinnamic acid from 4-nitrocinnamic acid, comprising:

using a second enzyme which consists of an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence defined in SEQ ID NOs:7, 9, 11, 13, or 15 and which has the ability to convert 4-nitrocinnamic acid to 4-aminocinnamic acid.

17. The method according to claim 10 , wherein the resting bacterial cell is selected from the group consisting of cultured cells, powdered cells, and immobilized cells.