Omposition of Reaction Item Under the above optimal circumstances, the reaction was performed as well as the composition with the reaction mixture was analyzed. The results are listed in Table four. Inside the original Pc, only compact amounts of 1-LPC and 2-LPC exist but 97.7 of pure Pc (Table 4). It was found that higher content of 1-LPC and 2-LPC was developed after 24 h of reaction, whereas the Computer content decreased to become 16.5 , and GPC emerged and accounted for 25.8 of total contents of reaction mixture. As shown in Table five, the DHA and EPA had been successfully incorporated into Computer and LPC. The content material of DHA and EPA inside the structured Pc and LPC was 30.8 and 30.3 , respectively. Table four. Composition of phospholipids of the original Pc and structured Pc.Phospholipids Computer 1-LPC 2-LPC GPCiContent ( ) Original Computer Structured Pc i 97.7 0.45 16.5 0.65 0.4 0.12 26.three 0.58 1.9 0.22 31.four 0.89 25.8 0.Reaction situations: enzyme loading, 15 (w/w); substrate mass ratio (PC/ethyl esters), 1:six; temperature, 55 ; water dosage, 1.25 (w/w); reaction time, 24 h; ” signifies not detected.two.four. Evaluation of Enzyme Reusability The reusability in the immobilized enzyme was evaluated below the optimal reaction conditions. The activity from the enzyme was calculated on the basis of your incorporation accomplished in 24 h reaction for each and every circle. The activity of immobilized PLA1 decreased to 48.9 after the fifth cycle (after it had been employed to become carried out a total of six reactions trials) (Figure 7). It indicated that immobilizedInt. J. Mol. Sci. 2014,PLA1 had a good operational stability in the transesterification of Computer and ethyl ester-DHA/EPA. Additionally, the immobilization technique applied inside the present work was incredibly easy and cost-effective, so the immobilized PLA1 has fantastic possible inside the production of structured phospholipids. Table five. Fatty acid composition ( ) of substrate and structured Computer.Fatty Acid C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:three C22:5 EPA DHA EPA + DHA Other people Substrate DHA/EPA-Rich Ethyl Esters 0.2 0.04 1.1 0.03 0.two 0.02 0.6 0.06 two.two 0.08 0.six 0.03 1.4 0.08 six.4 0.12 38.1 0.24 45.five 0.31 83.6 0.32 3.7 0.11 Computer k 1.1 0.05 13.8 0.13 0.three 0.01 three.six 0.08 10.6 0.18 64.8 0.32 five.8 0.07 LPC k 1.0 0.04 13.3 0.15 0.2 0.04 3.7 0.ten 11.1 0.25 65.2 0.27 5.5 0.BET bromodomain inhibitor 11 Structured Computer j Total Pc l Total LPC m 0.five 0.BT424 01 0.PMID:25147652 4 0.03 7.1 0.25 8.0 0.22 0.three 0.04 0.two 0.03 2.1 0.14 2.4 0.08 8.4 0.23 eight.6 0.12 43.1 0.35 42.9 0.31 3.eight 0.16 three.six 0.08 two.five 0.18 2.3 0.14 15.six 0.25 15.two 0.22 15.two 0.22 15.1 0.19 30.8 0.19 30.three 0.23 1.4 0.10 1.three 0.The original Pc was composed of 97.7 Pc and 2.3 LPC; j Reaction conditions: enzyme loading, 15 (w/w); substrate mass ratio (PC/ethyl esters), 1:six; temperature, 55 ; water dosage, 1.25 (w/w); reaction time, 24 h; k Computer and k LPC: the original unmodified Computer and LPC; l Total Pc: unreacted and modified Pc; m Total LPC: unreacted LPC that was present inside the original reaction mixture plus the LPC formed by hydrolysis and esterification; ” suggests not detected.Figure 7. Evaluation of immobilized PLA1 reusability. Reaction circumstances: enzyme loading, 15 (w/w); substrate mass ratio (PC/ethyl esters), 1:6; temperature, 55 ; water dosage, 1.25 (w/w); reaction time, 24 h.100Residual activity ( )80 70 60 50 40 30 20 0 1 2 three 4Recycle numberInt. J. Mol. Sci. 2014, 15 three. Experimental Section three.1. MaterialsGranulated soy phosphatidylcholine (Computer, purity 95 , utilised as reaction substrate) and 1-palmitoylsn-glycero-3-lysophosphatidylcholine (2-LPC, purity 99 ) were obtained from.