Patent ID: 8357503

Claim:
A method of hydrolyzing an oil or fat comprising reacting the oil or fat with a palmitase enzyme in presence of an emulsifier having HLB greater than 12, wherein palmitase enzyme is encoded by a nucleic acid sequence having at least 85% sequence identity to SEQ ID NO:1 and having i) nucleotide changes (or the equivalent thereof) encoding the amino acid residues at positions 85 and 172 (or the equivalent thereof) recited in Table A below: TABLE A 48 85 95 116 144 A48C E116I A144I A48C E116I A48C R85Y A48C R85Y E95K E116L A48C R85Y E95K A48C R85Y E95K A48C R85Y E116I R85Y E116L R85Y E95K E116L A48C E95K A48C R85Y E116L E95K R85Y A48C R85Y E95K E116L A48C R85Y E116I A144I A48C R85Y E116L A48C E116L A48C R85Y E95K E116L R85Y E95K R85Y A48C R85Y A48C R85Y E95K E116N A144I A48C E95K E116N A144I E116N R85Y E116I A48C R85Y E116L A48C E116I R85Y A48C R85Y A48C E116I A48C R85Y E116N A48C E116L E116I R85Y A48C R85Y E95K E116I R85Y E95K E116I R85Y E95K E95K E95K E116I R85Y E95K A48C E116L A48C R85Y E116N A144I A48C E95K E116I A144I E116I A144I R85Y A144I A48C E116N A144I A48C E116N A144I A48C R85Y E116I R85Y E116N 149 150 162 172 225 E149H A150I R172H A225S R172H A225S R172H R172H A225S P162G R172H A225S P162G R172H P162K R172H A225S P162G A225S P162G R172H A225S R172H A225S A150I P162G R172H A225S P162G R172H A225S P162G R172H A225S P162G R172H A225S R172H A225S P162K A150I R172H A225S P162G R172H A225S P162G R172H A225S P162G R172H A225S P162G R172H P162G R172H A225S P162K R172H P162K R172H P162K R172H A225S A150I P162G R172H P162G R172H P162G R172H P162G R172L A225S P162G R172H A225S P162G R172L A225S P162G R172L P162G P162G R172H A225S P162G P162G R172L A225S P162G A225S P162G P162G R172H P162G A225S P162G R172H P162G R172H P162G R172L A225S P162G R172L R172H A225S R172H A225S E149H R172H A225S R172H A150I A150I R172H E149H A150I R172H R172H ii) a nucleotide change (or the equivalent thereof) encoding the amino acid residue at position 83 (or the equivalent thereof) recited in Table B below: TABLE B old new Amino old new amino amino acid codon codon acid acid position GTG ATG V M 83 and iii) the following silent mutations 35GCT, 102GTT, 108AGT, 117CTT, 126AGG, 133TCT, and 188ACG.