Example 24
Stereospecific analysis of fatty acid species at sn-2 position of TAG from engineered seeds was conducted to assess the efficiency of assembly of short and medium chain fatty acids in TAG. Fatty acid profile of sn-2 monoacylglycerol obtained by digesting with TAG sn-1 and sn-3 specific lipase from Rhizomucor miehei (Sigma) revealed that while trace amounts of 10:0 is seen at sn-2 position of TAG from seeds of CvFatB1+CpDGAT1 lines, there is a striking increase up to 20 mol % in CvFatB1+CvLPAT2 line and 33.1 mol % in the relative content of 10:0 at sn-2 of TAG from CvFatB1+CvLPAT2+CpDGAT1 line was observed (
The increase of 10:0 at the sn-2 position of TAG from CvFatB1 expressing lines is accompanied by reduction of 18:2 and significantly that of 18:3 which is 17.4 mol % as compared to 44.3 mol % at this position in TAG from Wt camelina seeds. In C. pulcherrima fatty acid species at sn-2 position of TAG are 8:0 (up to 97 mol %) and C10:0 (3 mol %) while in C. viscosissima it is 12.6 mol % of C8:0 and 87.4 mol % of 10:0 (