Lipolytic enzymes

摘要

The inventors have developed a method using protein engineering to produce lipolytic enzymes having a relatively high activity for one ester bond in an amphiphilic substrate with two lipophilic groups) and a relatively low activity for the ester bond in an amphiphilic substrate with one lipophilic group, e.g. a relatively high phospholipase activity and a relatively low lysophospholipase activity.

主权项

1. An isolated lipolytic enzyme variant, comprising a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO: 14 and comprises an amino acid substitution R84G, R84A, R84Y, R84S, or L206F of SEQ ID NO: 14, and wherein the variant has phospholipase activity and lysophospholipase activity, and has a lysophospholipase to phospholipase activity ratio (PLARN) below 1000, wherein phospholipase activity is determined at 30° C. using 4% (w/v) lecithin (phosphatidyl choline) in 50 mM sodium acetate, 5 mM CaCl2, pH 5.0, and lysophospholipase activity is determined at 30° C. using 1% (w/v) lysolechitin in 50 mM sodium acetate, 5 mM CaCl2, pH 5.0.