Macrocyclic cyclases of peptides

Abstract

A peptide cyclase having the amino acid sequence shown in SEQ ID NO: 1 or a mutated sequence thereof, or a peptide cyclase having an amino acid sequence encoded by a base sequence encoding the amino acid sequence shown in SEQ ID NO: 1 or a mutated sequence thereof, DNA encoding the peptide cyclase, a method for producing the peptide cyclase, and a method for producing a cyclic peptide using the peptide cyclase.

Claims

1. (a) Sequence ID: The amino acid sequence shown in 1, (b) An amino acid sequence having 35% or more identity to the amino acid sequence shown in SEQ ID NO: 1, wherein the amino acid sequence of the portion corresponding to amino acid residues 63 to 66 of SEQ ID NO: 1 is Ser-X １ -X ２ -Lys, and the amino acid sequence of the portion corresponding to amino acid residues 153 to 158 of SEQ ID NO: 1 is Ser-Tyr-Ser-Asn-X ３ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 304 to 307 of SEQ ID NO: 1 is Gly-His-X ４ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 374 to 379 of SEQ ID NO: 1 is Gly-X ５ -X ６ -X ７ -Asn-Gly, where X １ ~X ７ each independently represents an amino acid residue, and the corresponding portion does not necessarily have to be the portion where the amino acid residue numbers of SEQ ID NO: 1 match, and may be a portion in its vicinity, or (c) An amino acid sequence in which one to ninety amino acid residues are deleted, substituted, inserted, or added in the amino acid sequence shown in SEQ ID NO: 1, wherein the amino acid sequence of the portion corresponding to amino acid residues 63 to 66 of SEQ ID NO: 1 is Ser-X １ -X ２ -Lys, and the amino acid sequence of the portion corresponding to amino acid residues 153-158 of SEQ ID NO: 1 is Ser-Tyr-Ser-Asn-X ３ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 304-307 of SEQ ID NO: 1 is Gly-His-X ４ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 374-379 of SEQ ID NO: 1 is Gly-X ５ -X ６ -X ７ -Asn-Gly, where X １ ~X ７ Each of these independently represents an amino acid residue, and the corresponding portion does not necessarily have to be the same as the amino acid residue number in SEQ ID NO: 1; it may be a nearby portion. A method for producing a cyclic peptide by head-to-tail cyclization, comprising reacting a substrate peptide with a peptide cyclase containing [a specific enzyme].

2. (a) The base sequence shown in Sequence ID No. 2, or (b) Sequence ID: A sequence that hybridizes under stringent conditions to a complementary sequence to the sequence shown in 2. A method for producing a cyclic peptide by head-to-tail cyclization, comprising acting a substrate peptide with a peptide cyclase containing an amino acid sequence encoded by DNA containing [specific DNA components].

3. For producing cyclic peptides by cyclization carried out in a head-to-tail manner, (a) Sequence ID: The amino acid sequence shown in 1, (b) An amino acid sequence having 35% or more identity with the amino acid sequence shown in SEQ ID NO: 1, wherein the amino acid sequence of the portion corresponding to amino acid residues 63 to 66 of SEQ ID NO: 1 is Ser-X １ -X ２ -Lys, and the amino acid sequence of the portion corresponding to amino acid residues 153-158 of SEQ ID NO: 1 is Ser-Tyr-Ser-Asn-X ３ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 304-307 of SEQ ID NO: 1 is Gly-His-X ４ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 374-379 of SEQ ID NO: 1 is Gly-X ５ -X ６ -X ７ -Asn-Gly, where X １ ~X ７ Each of these independently represents an amino acid residue, and the corresponding portion does not necessarily have to be the portion where the amino acid residue number matches that of SEQ ID NO: 1; it may be a nearby portion, or (c) An amino acid sequence in which one to ninety amino acid residues are deleted, substituted, inserted, or added in the amino acid sequence shown in SEQ ID NO: 1, wherein the amino acid sequence of the portion corresponding to amino acid residues 63 to 66 of SEQ ID NO: 1 is Ser-X １ -X ２ -Lys, and the amino acid sequence of the portion corresponding to amino acid residues 153-158 of SEQ ID NO: 1 is Ser-Tyr-Ser-Asn-X ３ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 304-307 of SEQ ID NO: 1 is Gly-His-X ４ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 374-379 of SEQ ID NO: 1 is Gly-X ５ -X ６ -X ７ -Asn-Gly, where X １ ~X ７ Each of these independently represents an amino acid residue, and the corresponding portion does not necessarily have to be the same as the amino acid residue number in SEQ ID NO: 1; it may be a nearby portion. Use of peptide cyclases containing [specific enzymes].

4. For producing cyclic peptides by cyclization carried out in a head-to-tail manner, (a) Sequence ID: A base sequence encoding the amino acid sequence shown in 1, (b) A base sequence encoding an amino acid sequence that has 35% or more identity with the amino acid sequence shown in SEQ ID NO: 1, wherein the amino acid sequence of the portion corresponding to amino acid residues 63 to 66 of SEQ ID NO: 1 is Ser-X １ -X ２ -Lys, and the amino acid sequence of the portion corresponding to amino acid residues 153-158 of SEQ ID NO: 1 is Ser-Tyr-Ser-Asn-X ３ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 304-307 of SEQ ID NO: 1 is Gly-His-X ４ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 374-379 of SEQ ID NO: 1 is Gly-X ５ -X ６ -X ７ -Asn-Gly, where X １ ~X ７ Each of these independently represents an amino acid residue, and the corresponding portion does not necessarily have to be the portion where the amino acid residue number matches that of SEQ ID NO: 1; it may be a nearby portion, or (c) A nucleotide sequence encoding an amino acid sequence in which one to ninety amino acid residues are deleted, substituted, inserted, or added in the amino acid sequence shown in SEQ ID NO: 1, wherein the amino acid sequence of the portion corresponding to amino acid residues 63 to 66 of SEQ ID NO: 1 is Ser-X １ -X ２ -Lys, and the amino acid sequence of the portion corresponding to amino acid residues 153-158 of SEQ ID NO: 1 is Ser-Tyr-Ser-Asn-X ３ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 304-307 of SEQ ID NO: 1 is Gly-His-X ４ -Gly, and the amino acid sequence of the portion corresponding to amino acid residues 374-379 of SEQ ID NO: 1 is Gly-X ５ -X ６ -X ７ -Asn-Gly, where X １ ~X ７ Each of these independently represents an amino acid residue, and the corresponding portion does not necessarily have to be the portion whose amino acid residue number matches that of Sequence ID No. 1; it may be a nearby portion. and the base sequence encoding the peptide cyclase Use of DNA containing [unclear].

5. For producing cyclic peptides by cyclization carried out in a head-to-tail manner, (a) The base sequence shown in Sequence ID No. 2, or (b) Sequence ID: A sequence that hybridizes under stringent conditions to a sequence complementary to the sequence shown in 2, and which encodes a peptide cyclase. Use of DNA containing [unclear].

6. For producing cyclic peptides by cyclization carried out in a head-to-tail manner, Use of a vector comprising DNA as described in claim 4 or 5.

7. A method for producing a cyclic peptide by cyclization in a head-to-tail manner, comprising reacting a substrate peptide with a peptide cyclase containing the amino acid sequence shown in SEQ ID NO: 5 or the amino acid sequence shown in SEQ ID NO:

9.

8. For producing cyclic peptides by cyclization carried out in a head-to-tail manner, The amino acid sequence shown in SEQ ID NO: 5 or the amino acid sequence shown in SEQ ID NO: 9 Use of peptide cyclases containing [specific enzymes].

Images