Enzymatic acyl modification of phosphatidylcholine using immobilized lipase and phospholipase A₂

Abstract

Ethanol-soluble (ES) lecithin mainly contains phosphatidylcholine (PC). The incorporation of caprylic acid into PC using immobilized phospholipase A 2 (PLA 2 ) and lipase was investigated. The Rhizomucor meihei lipase and the porcine pancreatic PLA 2 were immobilized on the hydrophobic resin Diaion HP-20 and the modification was carried out in hexane as solvent. HPTLC with densitometer technique was successfully used for monitoring the production of structured phospholipids (PL) (ML-type PC, MM-type PC, and lysophosphatidylcholine; L: long-chain fatty acid, M: medium-chain fatty acid). The various parameters such as the effects of reaction temperature, enzyme loading, and the effect of molar proportion of substrate were studied in order to determine the optimum reaction conditions for the acidolysis reaction. The optimal operating conditions for the PLA 2 -catalyzed reaction were obtained as 508C temperature, 50% (wt/wt of substrate) enzyme loading, and a 1:12 molar proportion of PC/caprylic acid. For the lipase-catalyzed reaction, the optimized temperature was the same as for PLA 2 , but the enzyme loading and molar proportion were slightly lower, i.e., 40 % w/w of substrate and 1:9 PC/caprylic acid, respectively. The effects of these parameters on the production of structured PL were compared. Under these optimal conditions, the ML-type PC content was higher in the PLA 2 -catalyzed reaction, i.e., 45.29 mol%, and in the lipase-catalyzed reaction it was 38.74 mol%.

A Short Review on Various Screening Methods to Isolate Potential Lipase Producers: Lipases-the Present and Future Enzymes of Biotech Industry

“…Released free fatty acids can also be quantitated by using High Performance Thin Layer Chromatography (HPTLC), a method that is more simple, precise, specific and accurate. Quantification of individual phospholipid compounds was performed by HPTLC in studies by More and Pandit (2010). The method was also used by Vitro et al (2000) for determining the fatty acid distribution.…”

Quantitative Lipidome Analysis of Boiled Chicken Egg Yolk under Different Heating Intensities

“…Egg yolks are rich in endogenous lipases, including acid lipase, neutral lipase, and phospholipase [ 37 , 38 ]. Among them, the optimum temperature of egg yolk phospholipase is 40–55 °C [ 39 , 40 ]. Therefore, the phospholipid hydrolysis may be accelerated in the temperature ramp-up period during the heating process, which is more likely to occur in HEY and SEY with a lower heating intensity.…”

Progress & Prospect of Enzyme-Mediated Structured Phospholipids Preparation

“…PLA2 (EC 3.1.1.4) can catalyze the exchange of fatty acid in the sn-2 position via acidolysis or the transesterification reaction [37,65] (Figure 8). Compared with a lipase-and PLA1-catalyzed reaction, PLA2-catalyzed acyl exchange has received little attention as PLA2 may lack the potential to catalyze the transesterification reaction since no acyl-enzyme intermediate is formed [66]. PLA 2 (EC 3.1.1.4) can catalyze the exchange of fatty acid in the sn-2 position via acidolysis or the transesterification reaction [37,65] (Figure 8).…”