Patent ID: 8349578

Claim:
A method for biocatalytic synthesis of a structured lipid comprising the following steps: (a) providing a polypeptide having a palmitase activity, said polypeptide being selected from the group consisting of isolated, synthetic and recombinant polypeptides having a palmitase activity and said polypeptide either i. being encoded by a nucleic acid comprising a nucleic acid sequence having at least 85% sequence identity to full length SEQ ID NO:1, and wherein the nucleic acid encodes at least one polypeptide having palmitase activity and the polypeptide having at least one amino acid residue change recited below in Table A, or ii. having at least 85% sequence identity to full length SEQ ID NO:2, and having at least one amino acid residue change recited in Table A, TABLE A Amino Original New Acid Residue Amino Acid Amino Acid 27 G Q 45 G L 54 S H 61 D A E 62 V A G 72 R E K S T 74 F I 83 V M 96 A I 98 G L 116 E A F H L Q Q R, H140R R S S T V 117 L R, I161L 120 K L 133 S A 147 I F 151 I A G H 160 P T 172 R Q 212 S R 213 K T 214 T V N R Y 215 G A H 216 A V C L or iii. having at least 85% sequence identity to full length of SEQ ID NO:2 and comprising at least one amino acid residue modification D61A; D61E; R72E; R72K; E116A; E116Q; E116R; E116T; E116V; S133A; I151G; I151A; V163R; or D164R (b) providing a composition comprising a triacylglyceride; (c) contacting the polypeptide of step (a) with the composition of step (b) under conditions wherein the polypeptide hydrolyzes a palmitic acid residue at the Sn1 or Sn3 position of the triacylglyceride, thereby producing a 1,2-diacylglyceride or 2,3-diacylglyceride; and (d) promoting acyl migration in the 1,2-diacylglyceride or 2,3-diacylglyceride of the step (c) under kinetically controlled conditions, thereby producing a 1,3-diacylglyceride.