Patent EP-4067497-A1 (EP)
Inventor: BABA Yutaro
IPC: C12N1/19
Filed: 2020-11-27
Abstract: An object of the present invention is to provide a technique relating to a method for enzymatically producing maytansinol from an ansamitocin species such as AP3. A method for producing maytansinol, which comprises enzymatically producing maytansinol from an ansamitocin species with any one of the proteins (A) to (F) and (AA) to (CC) mentioned below: (A) a protein consisting of the amino acid sequence of SEQ ID NO: 9; (B) a protein consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 9 by substitution, deletion, insertion, and/or addition of one or more amino acids, wherein amino acids corresponding to the positions 66 to 68 are DLL or AGA, amino acids corresponding to the positions 139 to 141 are GPF, amino acids corresponding to the positions 146 to 148 are LSS or LSP, amino acids corresponding to the positions 247 to 249 are LHT, amino acids corresponding to the position 323 are HSX or ASX (where X is Q, K, E, or M), amino acids corresponding to the positions 374 to 376 are QSX (where X is H, R, D, or Q), and amino acids corresponding to the positions 443 to 445 are GNP, and having an activity for catalyzing a reaction of producing maytansinol from an ansamitocin species; (C) a protein consisting of an amino acid sequence having an identity of at least 50% to the amino acid sequence of SEQ ID NO: 9, wherein amino acids corresponding to the positions 66 to 68 are DLL or AGA, amino acids corresponding to the positions 139 to 141 are GPF, amino acids corresponding to the positions 146 to 148 are LSS or LSP, amino acids corresponding to the positions 247 to 249 are LHT, amino acids corresponding to the position 323 are HSX or ASX (where X is Q, K, E, or M), amino acids corresponding to the positions 374 to 376 are QSX (where X is H, R, D, or Q), and amino acids corresponding to the positions 443 to 445 are GNP, and having an activity for catalyzing a reaction of producing maytansinol from an ansamitocin species; (D) a protein consisting of the amino acid sequence of SEQ ID NO: 12; (E) a protein consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 12 by substitution, deletion, insertion, and/or addition of one or more amino acids, wherein amino acids corresponding to the positions 66 to 68 are DLL or AGA, amino acids corresponding to the positions 139 to 141 are GPF, amino acids corresponding to the positions 146 to 148 are LSS or LSP, amino acids corresponding to the positions 247 to 249 are LHT, amino acids corresponding to the position 323 are HSX or ASX (where X is Q, K, E, or M), amino acids corresponding to the positions 374 to 376 are QSX (where X is H, R, D, or Q), and amino acids corresponding to the positions 443 to 445 are GNP, and having an activity for catalyzing a reaction of producing maytansinol from an ansamitocin species; (F) a protein consisting of an amino acid sequence having an identity of at least 50% to the amino acid sequence of SEQ ID NO: 12, wherein amino acids corresponding to the positions 66 to 68 are DLL or AGA, amino acids corresponding to the positions 139 to 141 are GPF, amino acids corresponding to the positions 146 to 148 are LSS or LSP, amino acids corresponding to the positions 247 to 249 are LHT, amino acids corresponding to the position 323 are HSX or ASX (where X is Q, K, E, or M), amino acids corresponding to the positions 374 to 376 are QSX (where X is H, R, D, or Q), and amino acids corresponding to the positions 443 to 445 are GNP, and having an activity for catalyzing a reaction of producing maytansinol from an ansamitocin species; (AA) a protein consisting of any one of the amino acid sequences of SEQ ID NOS: 18 to 21; (BB) a protein consisting of an amino acid sequence derived from any one of the amino acid sequences of SEQ ID NOS: 18 to 21 by substitution, deletion, insertion, and/or addition of one or more amino acids, and having an activity for catalyzing a reaction of producing maytansinol from an ansamitocin species; and (CC) a protein consisting of an amino acid sequence having an identity of at least 80% to any one of the amino acid sequences of SEQ ID NOS: 18 to 21, and having an activity for catalyzing a reaction of producing maytansinol from an ansamitocin species.