Production of Structured Phosphatidylcholine with High Content of Myristic Acid by Lipase-Catalyzed Acidolysis and Interesterification

Abstract

Synthesis of structured phosphatidylcholine (PC) enriched with myristic acid (MA) was conducted by acidolysis and interesterification reactions using immobilized lipases as catalysts and two acyl donors: trimyristin (TMA) isolated from ground nutmeg, and myristic acid obtained by saponification of TMA. Screening experiments indicated that the most effective biocatalyst for interesterification was Rhizomucor miehei lipase (RML), whereas for acidolysis, the most active were Thermomyces lanuginosus lipase (TLL) and RML. The effect of the molar ratio of substrates (egg-yolk PC/acyl donor), enzyme loading, and different solvent on the incorporation of MA into PC and on PC recovery was studied. The maximal incorporation of MA (44 wt%) was achieved after 48 h of RML-catalyzed interesterification in hexane using substrates molar ratio (PC/trimyristin) 1/5 and 30% enzyme load. Comparable results were obtained in toluene with 1/3 substrates molar ratio. Interesterification of PC with trimyristin resulted in significantly higher MA incorporation than acidolysis with myristic acid, particularly in the reactions catalyzed by RML.

Novozym 435: the “perfect” lipase immobilized biocatalyst?

“…42 The heterogeneity of the natural substrate [43][44][45][46][47][48] has converted lipases in enzymes with a very broad specificity, accepting substrates very different from glycerides (even amides). Thus, lipases are used in vitro to catalyze reactions different from those of the natural hydrolase function, [49][50][51][52][53][54] such as esterification, [55][56][57][58][59][60][61] acidolysis, [62][63][64][65][66][67] interesterificaton, [68][69][70][71] transesterification, [72][73][74][75][76] aminolysis, [77][78][79][80][81] perhydrolysis, 82,83 etc., together with a collection of the so-called promiscuous reactions. [84][85][86][87][88]...…”

Lecitase ultra: A phospholipase with great potential in biocatalysis

“…Among the most used enzymes, lipases stand out [42,43]. The natural function of lipases is the hydrolysis of triglycerides to free fatty acids and glycerol [44]; however, in vitro lipases have been used in a wide variety of reactions, some related to oil modifications (esterifications [45][46][47][48][49][50][51], transesterifications [52][53][54][55][56], acidolysis [57][58][59][60][61][62], interesterifcations [63][64][65][66]), some far from these substrates (resolution of racemic mixtures, regio or enantio selective reactions) and some have even exhibited a diversity of the so-called promiscuous activities far from their natural function [67][68][69][70][71][72][73][74][75].…”

Lipase Catalyzed Acidolysis for Efficient Synthesis of Phospholipids Enriched with Isomerically Pure cis-9,trans-11 and trans-10,cis-12 Conjugated Linoleic Acid

“…No significant differences in INCLPC was observed between the isomers in case of aw = 0.11 and 0.23. The obtained results are consistent with the literature data, which 14 22.3 ± 0.9 18 14.4 ± 3.7 23 0.33 6.4 ± 0.1 4 24.4 ± 0.8 9 11.8 ± 0.3 6 42.0 ± 0.8 7 36.1 ± 0.5 12 15.5 ± 0.8 16 48.0 ± 0.8 22 18.3 ± 0.8 15 8.7 ± 0.3 19 73.6 ± 3.0 24 1 Results refers to 48 h of the acidolysis reaction. condition are given in Figure 5.…”

“…Vertical bars represent SD, n = 3. 3 22.7 ± 3.9 7 2.6 ± 0.4 5 3.4 ± 0.4 10 2.6 ± 0.4 5 3.4 ± 0.4 10 38.2 ± 3.4 11,12 11.7 ± 0.6 20 50.1 ± 4.0 19 53.2 ± 1.0 14 30.9 ± 1.2 18 15.9 ± 2.2 22 TLL 5.9 ± 0.4 4 5.3 ± 0.5 8 3.1 ± 0.6 3,5 nd 37.1 ± 0.4 12 13.9 ± 0.3 17 49.0 ± 0.7 19 42.7 ± 0.5 15 30.6 ± 1.1 18 26.7 ± 1.5 23 1 Results refers to 24 h of the acidolysis reaction, condition are given in Figure 2. The values are mean ± SD based on three independent experiments.…”

“…Naturally occurring glycerophospholipids usually composed of saturated fatty acids, such as stearic or palmitic acid in the sn-1 position, while the sn-2 position is occupied by unsaturated acids (i.e., oleic, linoleic, and arachidonic) [14]. Structured phospholipids are the products of such modifications, and the process itself involves rearrangement, addition or replacement of fatty acid residues in the sn-1 or sn-2 position of glycerol backbone with desired fatty acids, for example medium chain saturated acids, to ensure high oxidative stability [15][16][17], polyunsaturated fatty acids (eicosapentaenoic and docosahexaenoic) that possess health-promoting properties [18][19][20] or conjugated fatty acids, such as conjugated linoleic acid (CLA) (Table 1). Amongst the PL modification processes, enzymatic methods are currently becoming the most popular because of many advantages (i.e., regioselectivity, substrate specificity, low temperature, and lack of harsh and harmful chemical reagents).…”