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1. WO2011000121 - POLYPEPTIDES HAVING TYROSINASE ACTIVITY AND USES THEREOF

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[ EN ]

Claims

1. A polypeptide being a fragment of the V. spinosum tyrosinase or a variant thereof wherein the fragment comprises the mature domain of the V. spinosum tyrosinase or a functional fragment thereof.

2. The polypeptide of claim 1, wherein the variant has at least 50% sequence identity, in particular at least 60%, 70%, 80%, 85%, 90%, 95% sequence identity, to SEQ. ID NO. 3.

3. The polypeptide of claim 1 or 2, comprising or consisting of SEQ ID No. 7.

4. The polypeptide of anyone of the preceding claims, wherein the fragment further comprises a signal peptide at its N-terminal end, in particular a signal peptide other than SEQ ID No. 4.

5. The polypeptide of anyone of the preceding claims, wherein the fragment further comprising a N- or C-terminal tag, in particular a C-terminal tag other than SEQ ID NO. 5.

6. A polypeptide comprising a sequence having at least 50% identity, in particular at least 60%, 70%, 80%, 85%, 90%, 95% sequence identity, to SEQ ID No. 6 with the proviso that the amino acid at position 453 is not phenylalanine, preferably is neither phenylalanine nor tyrosine nor leucine, most preferably the amino acid at position 453 is alanine.

7. The polypeptide of claim 6, having SEQ ID No. 6.

8. An isolated nucleic acid encoding a fragment of the V. spinosum tyrosinase or a variant thereof and optionally one or two adjacent polypeptide sequences other than SEQ ID No. 4 and/or SEQ. ID. No. 5.

9. The isolated nucleic acid of claim 8, wherein the fragment is the polypeptide of anyone of claims 1 to 7.

10. The isolated nucleic acid of claim 8 or

9, comprising SEQ ID No. 8 or a fragment thereof; or being capable of hybridizing with a sequence comprised in SEQ ID No. 8.

11. A vector comprising the nucleic acid of claims 8 to 10.

12. A vector comprising a nucleic acid encoding a polypeptide at least 75% identical to SEQ ID No: 1, wherein the polypeptide has tyrosinase activity.

13. A host cell comprising the vector of anyone of claims 11 or 12.

14 . Use of the polypeptide of anyone of claims 1-7 in the synthesis of melanin.

15. Use of the polypeptide of anyone of claims 1-7 in the crosslinking of tyrosine-containing proteins .

16. The use according to claim 15, wherein the protein is a food protein, a proteinaceous fiber or a fiber-derived polymer.

17. A method for the production of melanin comprising the steps of

(a) providing L-tyrosine

(b) providing the tyrosinase of anyone of claims 1-7; and

(c) reacting the L-tyrosine with the tyrosi-nase under oxidizing conditions.

18. A method for the cross-linking of proteins comprising the steps of

(a) providing a first polypeptide comprising tyrosine at one or more positions;

(b) optionally providing a second polypeptide comprising tyrosine at one or more positions;

(c) providing the tyrosinase of anyone of claims 1-7; and

(d) reacting the first and optionally the second polypeptide with the tyrosinase under oxidizing conditions .

19. A method for the cross-linking of phenols comprising the steps of

(a) providing mono- and/or bisphenols;

(b) providing one or more polymers having amino-, SH-, imidazol- and/or phenylgroups as side chains;

(c) providing the tyrosinase of anyone of claims 1-7; and

(d) reacting the phenols with the polymers and the tyrosinase under oxidizing conditions.