Surfactant tolerant cellulase and method for modification thereof
A method for suppressing a reduction in an endoglucanase activity in the presence of a surfactant, characterized by modifying a protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid, to a protein having the N-terminus of pyroglutamic acid, is disclosed. Further, a modified protein having an endoglucanase activity wherein the N-terminal amino acid is converted into pyroglutamic acid by an amino acid modification, a polynucleotide encoding the protein, an expression vector comprising the polynucleotide, a host cell transformed with the expression vector, and a process for producing the protein by cultivating the host cell, are disclosed.
1. An isolated protein selected from the group consisting of:
(a) a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 38, or 40, wherein the N-terminal amino acid is pyroglutamic acid; and
(b) a protein comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 2, 4, 38, or 40, and having an endoglucanase activity, wherein the N-terminal amino acid is pyroglutamic acid.
2. A protein produced by the steps comprising:
cultivating a host cell transformed with an expression vector comprising a polynucleotide encoding the protein according to claim 1 , and recovering the protein from the host cell or culture obtained by the cultivation.
3. A method for suppressing a reduction in an endoglucanase activity in the presence of a surfactant, characterized by modifying a protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid, to a protein according to claim 1 .
4. The method according to claim 3 , wherein the modification is carried out by adding pyroglutamic acid or an amino acid convertible into pyroglutamic acid, or a peptide having the N-terminus of pyroglutamic acid or an amino acid convertible into pyroglutamic acid, to the N-terminus of the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid.
5. The method according to claim 3 , wherein the modification is carried out by substituting pyroglutamic acid or an amino acid convertible into pyroglutamic acid, or a peptide having the N-terminus of pyroglutamic acid or an amino acid convertible into pyroglutamic acid, for the N-terminal amino acid or an N-terminal region of the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid.
6. The method according to claim 3 , wherein the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid is a cellulase belonging to family 45.
7. The method according to 4 , wherein the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid is a cellulase belonging to family 45.
8. The method according to claim 5 , wherein the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid is a cellulase belonging to family 45.