Patent ID: 9187738
Filing Date: 2015-11-17
Classification: A21D,C12N,C12Y

Abstract:
1. A method for preparing a variant lipolytic enzyme with improved lipase activity as compared with the precursor wild-type KLM1enzyme having the amino acid sequence SEQ ID NO: 2 comprising expressing in a host organism a nucleotide sequence encoding a variant lipolytic enzyme the precursor amino acid sequence of which has at least 85% identity with the amino acid sequence of the precursor lipase amino acid sequence from position 31 through position 348 of SEQ ID No. 2 wherein the encoded precursor variant lipolytic enzyme is modified such that one or more amino acids located at a surface position within an external loop distal to the active site of the variant precursor lipolytic enzyme is substituted with an amino more hydrophilic than the original amino acid (i) at a position corresponding to any of positions 54-66 of SEQ ID No. 2, (ii) at a position corresponding to any of positions 75-79 of SEQ ID No. 2, (iii) at a position corresponding to any of positions 99-103 of SEQ ID No. 2, (iv) at a position corresponding to any of positions 127-135 of SEQ ID No. 2, (v) at a position corresponding to any of positions 162-167 of SEQ ID No. 2, (vi) at a position corresponding to any of positions 188-195 of SEQ ID No. 2, and/or (vii) at a position corresponding to any of positions 213-221 of SEQ ID No. 2; wherein the modification is not a substitution at position 63.