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

Abstract

The production of phospholipid (PL) conjugates with biologically active compounds is nowadays an extensively employed approach. This type of phospholipids conjugates could improve bioavailability of many poorly absorbed active compounds such as isomers of conjugated linoleic acid (CLA), which exhibit versatile biological effects. The studies were carried out to elaborate an efficient enzymatic method for the synthesis of phospholipids with pure (>90%) cis-9,trans-11 and trans-10,cis-12 CLA isomers. For this purpose, three commercially available immobilized lipases were examined in respect to specificity towards CLA isomers in acidolysis of egg-yolk phosphatidylcholine (PC). Different incorporation rates were observed for the individual CLA isomers. Under optimal conditions: PC/CLA molar ratio 1:6; Rhizomucor miehei lipase loading 24% wt. based on substrates; heptane; DMF, 5% (v/v); water activity (aw), 0.11; 45 °C; magnetic stirring, 300 rpm; 48 h., effective incorporation (EINC) of CLA isomers into PC reached ca. 50%. The EINC of CLA isomers was elevated for 25–30% only by adding a water mimic (DMF) and reducing aw to 0.11 comparing to the reaction system performed at aw = 0.23. The developed method of phosphatidylcholine acidolysis is the first described in the literature dealing with isometrically pure CLA and allow to obtain very high effective incorporation.

Lipase-Catalyzed Preparation and Optimization of Structured Phosphatidylcholine Containing Nervonic Acid

“…Verdasco-Martín, et al [24] reported that using a PC sample without dehydration severely affected reaction efficiency caused by undesired hydrolysis and strongly competes with the desired esterification of FAs. This could be caused by existing water in the substrates or water produced in the reaction adequate to maintain lipase activity in the reaction [33,34]. Li, et al [35] reported that a hydrolysis reaction occurred even though the water addition was 0% wt which may be contributed by the inherent moisture of the immobilized PLA 1 that results in the continued decrease in PC recovery.…”

Progress & Prospect of Enzyme-Mediated Structured Phospholipids Preparation

“…On one hand, the presence of water in the reaction system is crucial for the hydrolysis of PLs and the activation of the enzyme. On another hand, under high content or activity of water, the additional moisture may either decrease the catalytic of the enzyme or lead to undesired hydrolysis reactions [30,63].…”

“…Levels of water addition also affected reaction time where the incorporation reached its maximum under a given water dosage and the final composition of the products. Niezgoda and Gliszczynska [30] investigated the effect of water activity on the efficiency of PC acidolysis with conjugated linoleic acid (CLA) under the condition of a w = 0.11, 0.23, and 0.33. They found that the effective incorporation of CLA declined in the series 0.23 > 0.33 > 0.11.…”

Thermally Stable and Reusable Silica and Nano-Fructosome Encapsulated CalB Enzyme Particles for Rapid Enzymatic Hydrolysis and Acylation

“…It has high activity and regioselectivity. Depending on reaction conditions, CalB can promote reactions such as hydrolysis and acidolysis [ 7 , 8 , 9 , 10 , 11 ]. CalB is expected to be a non-toxic and renewable energy source.…”