Patent ID: 7157262

Claim:
A method of preparing a mutated lipolytic enzyme, which method comprises: a) subjecting DNA sequences encoding parent lipolytic enzyme to mutagenesis to form a variety of mutated DNA sequences encoding mutated lipolytic enzymes having, as compared to the parent lipolytic enzyme, a peptide addition at the N-terminus, at the C-terminus or at both the N-terminus and C-terminus; b) expressing the mutated DNA sequences in host cells which secrete said mutated lipolytic enzymes having the peptide addition after transcription, translation and processing; c) selecting a mutated lipolytic enzyme among those resulting from step (b) which, when present in a detergent composition, is capable of removing at least 15% more lard from a lard stained swatch, than the same detergent composition without the lipolytic enzyme, in a one cycle wash assay comprising subjecting 7 lard-stained cotton swatches (9×9 cm) per beaker to a one cycle wash in a thermostated TOM, each beaker containing 1000 ml of water comprising 3.2 mM Ca2+/Mg2+ (in a ratio of 5:1) and 5 g/l of said detergent composition, adjusted to pH 10, and comprising 12500 LU/I of the lipolytic enzyme, for 20 minutes at a temperature of 30° C., followed by rinsing for 15 minutes in running tap water and overnight linedrying at room temperature, subsequent extraction and quantification of fatty matter from the resulting swatches by Soxhlet extraction.