Fied reaction equation shown in Scheme 1), and reaction situations, for example enzyme load, substrates mass ratio, reaction temperature and water dosage, have been investigated. Scheme 1. Transesterification reaction of Computer with DHA/EPA-rich ethyl esters.O O O O R1 C O CH2 PLA 1 EPA/DHA C O CH two R2 C O CH R2 C O CH O O CH 3 CH3 CH 2O P O CH2CH2 +N CH3 DHA/EPA O O OCH2CH3 CH 2 OP O CH2CH2 +N CH3 R1 OCH2CH3 CH3 CH3 OO-2.2.1. Impact of Enzyme Loading Effect of immobilized PLA1 loading around the transesterification reaction was evaluated. Enzyme loading is the percentage in the immobilized enzyme weight (containing enzyme itself and also the resin D380) for the total weight of reaction substrates (Pc and ethyl esters). The reaction was performed at 50 for 24 h with 1:4 of substrate mass ratio (Computer to DHA/EPA-rich ethyl esters) and 1 of water dosage. The enzyme loading was chosen as 5 , 10 , 15 , 20 , 25 (w/w, with respect to total reaction mixture), respectively.Meropenem The outcomes are shown in Figure 5. The incorporation of DHA and EPA into Computer enhanced together with the immobilized PLA1 load growing. When ten immobilized PLA1 wasInt. J. Mol. Sci. 2014,employed, the incorporation of DHA and EPA reached additional than 19 following 24 h, and were 19.5 and 23.eight respectively with the enzyme loading at 10 and 15 . The optimal enzyme loading was 15 , along with the incorporation of EPA and DHA had been 12.four and 11.four , respectively. Figure five. Effect of immobilized phospholipase A1 loading around the transesterification reaction. Reaction circumstances: substrate mass ratio (Computer: ethyl esters), 1:4; temperature, 50 ; water dosage, 1 (w/w); reaction time, 24 h.Total incorporation of EPA and DHA ( )30Enzyme loading ( )2.Dinutuximab two.PMID:24025603 2. Impact of Mass Ratio of Computer to DHA/EPA-Rich Ethyl Esters Effect of mass ratio of Pc to DHA/EPA-rich ethyl esters on the transesterification reaction was investigated. The reaction was performed at 50 with the enzyme loading of 15 . Plus the mass ratio of Pc to DHA/EPA-rich ethyl esters was set as 1:three, 1:four, 1:5, 1:six and 1:7, respectively. The outcomes are shown in Figure six. When the mass ratio of Computer to DHA/EPA-rich ethyl esters varied from 1:3 to 1:six, the incorporation of DHA and EPA increased and it accomplished a maximum of 25.six at 1:6, whereas much less increment was observed at 1:7. The incorporation of EPA and DHA respectively reached the maximum of 13.0 and 12.six when substrate mass ratio was 1:6. Excess DHA/EPA-rich ethyl esters would advantage the incorporation of DHA/EPA into Computer. This may be explained that excess DHA/EPA-rich ethyl esters may raise the concentration of DHA/EPA, the solubility of Computer and decrease mass transfer. Therefore, the mass ratio was fixed at 1:six within the latter reaction program. two.2.three. Impact of Temperature Effect of temperature around the transesterification was explored. The temperature was set as 40, 45, 50, 55 and 60 , respectively. The enzyme loading was fixed at 15 , and mass ratio of Pc to DHA/EPA-rich ethyl esters was kept at 1:6. The outcomes are shown in Table 2. Compared using the initial ratio of EPA to DHA (0.84) within the DHA/EPA-rich ethyl esters, the ratio of EPA to DHA changed to become 1.two, 1.11, 1.04, 1.01, 0.98 after 24 h of reaction at 40, 45, 50, 55, 60 respectively. It was indicated that immobilized PLA1 showed slightly much more preference to EPA than DHA whereas the tendency decreased using the reaction temperature rising. The incorporation of DHA/EPA into Pc enhanced obviously with temperature increasing and reached a maximum of 27.eight at 55 .