PDAT Activity and Substrate Specificity of Recombinant Cr-PDAT.
(A) PDAT activity of ORFCrPDAT and ΔTMCrPDAT. The sn-1,2-diolein (18:1 DAG) and different PLs, including PA, PS, PI, PG, PC, and PE, were used as substrates. Heat-inactivated PDAT was used as negative control (NC) for the assay. FFA, free fatty acid.
(B) Effects of different PL concentrations on the biosynthesis of TAG.
(C) Acyl donor side-chain specificity of Cr-PDAT. The substrate specificity was determined with PE containing various sn-2-acyl groups, and sn-1,2-diolein was used as an acyl acceptor.
(D) Acyl acceptor positional specificity of Cr-PDAT. The substrate specificity was determined with 1,2-dioleoyl-sn-glycerol (1,2-DAG) and 1,3-dioleoyl-sn-glycerol (1,3-DAG). Various PLs (PA, PC, PE, PS, PI, and PG) were used as acyl donors.
(E) Acyl acceptor side-chain specificity of Cr-PDAT. The substrate specificity was determined with 1,2-dipalmitoyl-sn-glycerol (16:0-DAG) and 1,2-dioleoyl-sn-glycerol (18:1-DAG). PI (250 µM) was used as an acyl donor. ΔTMCrPDAT was used in the enzymatic assays ([B] to [E]).