METHOD FOR DESIGNING MUTANT ENZYME, METHOD FOR PREPARING MUTANT ENZYME, AND MUTANT ENZYME

摘要

An object is to provide a novel method of improving an enzyme capable of deamidating a protein. A mutant enzyme is designed by the following steps: (1) specifying one or more amino acids selected from the following group, namely, consisting of an amino acid corresponding to the amino acid at position 35, an amino acid corresponding to the amino acid at position 38, an amino acid corresponding to the amino acid at position 39, an amino acid corresponding to the amino acid at position 40, an amino acid corresponding to the amino acid at position 41, an amino acid corresponding to the amino acid at position 42, an amino acid corresponding to the amino acid at position 43, an amino acid corresponding to the amino acid at position 45, an amino acid corresponding to the amino acid at position 46, an amino acid corresponding to the amino acid at position 49, an amino acid corresponding to the amino acid at position 79, an amino acid corresponding to the amino acid at position 80, an amino acid corresponding to the amino acid at position 81, an amino acid corresponding to the amino acid at position 82, an amino acid corresponding to the amino acid at position 83, an amino acid corresponding to the amino acid at position 84, an amino acid corresponding to the amino acid at position 103, an amino acid corresponding to the amino acid at position 104, an amino acid corresponding to the amino acid at position 105, an amino acid corresponding to the amino acid at position 106, an amino acid corresponding to the amino acid at position 117, an amino acid corresponding to the amino acid at position 142, an amino acid corresponding to the amino acid at position 143, an amino acid corresponding to the amino acid at position 146, an amino acid corresponding to the amino acid at position 166, and an amino acid corresponding to the amino acid at position 185 in an amino acid sequence set forth in SEQ ID NO: 2, in a protein deamidase (an enzyme to be mutated); and (2) constructing an amino acid sequence having substitution of the amino acid(s) specified in the step (1) by another amino acid(s) or having deletion of the amino acid(s) specified in the step (1) using the amino acid sequence for an enzyme to be mutated as a base sequence.