Patent WO-2021107133-A1 (WO)
Inventor: BABA Yutaro
IPC: C12N9/18
Filed: 2020-11-27
Abstract: The present invention addresses the problem of providing a technique related to a method for enzymatically producing maytansinol from ansamitocins such as AP3.　Provided is a method for producing maytansinol, the method including a step for enzymatically generating maytansinol from ansamitocins using one protein of (A)-(F) and (AA)-(CC): (A) a protein comprising the amino acid sequence represented by SEQ ID NO:9; (B) a protein comprising the amino acid sequence represented by SEQ ID NO: 9 with one or more amino acids substituted, deleted, inserted and/or added, where positions 66-68 correspond to residues DLL or AGA, positions 139-141 correspond to residues GPF, positions 146-148 correspond to residues LSS or LSP, positions 247-249 correspond to residues LHT, position 323 corresponds to residues HSX or ASX (where X is Q, K, E or M), positions 374-376 correspond to residues QSX (where X is H, R, D or Q), positions 443-445 correspond to residues GNP, and having activity for catalyzing a reaction that produces maytansinol from ansamitocins; (C) a protein having at least 50% amino acid identity with the amino acid sequence represented by SEQ ID NO: 9, where positions 66-68 correspond to residues DLL or AGA, positions 139-141 correspond to residues GPF, positions 146-148 correspond to residues LSS or LSP, positions 247-249 correspond to residues LHT, positions 323 correspond to residues HSX or ASX (where X is Q, K, E or M), positions 374-376 correspond to residues QSX (where X is H, R, D or Q), positions 443-445 correspond to residues GNP, and having activity for catalyzing a reaction that produces maytansinol from ansamitocins; (D) a protein comprising the amino acid sequence represented by SEQ ID NO: 12; (E) a protein comprising the amino acid sequence represented by SEQ ID NO: 12 with one or more amino acids substituted, deleted, inserted and/or added, where positions 66-68 correspond to residues DLL or AGA, positions 139-141 correspond to residues GPF, positions 146-148 correspond to residues LSS or LSP, positions 247-249 correspond to residues LHT, position 323 corresponds to residues HSX or ASX (where X is Q, K, E or M), positions 374-376 correspond to residues QSX (where X is H, R, D or Q), positions 443-445 correspond to residues GNP, and having activity for catalyzing a reaction that produces maytansinol from ansamitocins; (F) a protein having at least 50% amino acid identity with the amino acid sequence represented by SEQ ID NO: 12, where positions 66-68 correspond to residues DLL or AGA, positions 139-141 correspond to residues GPF, positions 146-148 correspond to residues LSS or LSP, positions 247-249 correspond to residues LHT, position 323 corresponds to residues HSX or ASX (where X is Q, K, E or M), positions 374-376 correspond to residues QSX (where X is H, R, D or Q), positions 443-445 correspond to residues GNP, and having activity for catalyzing a reaction that produces maytansinol from ansamitocins; (AA) a protein comprising the amino acid sequence represented by any one of SEQ ID NOs:18-21; (BB) a protein comprising the amino acid sequence represented by any one of SEQ ID NOs:18-21 with one or more amino acids substituted, deleted, inserted and/or added, and having activity for catalyzing a reaction that produces maytansinol from ansamitocins; and (CC) a protein having at least 80% amino acid identity with the amino acid sequence represented by any one of SEQ ID NOs:18-21, and having activity for catalyzing a reaction that produces maytansinol from ansamitocins.