Enzymatic Hydrolysis of Natural Lipids in Polymeric Membranes

Enzymatic Hydrolysis of Natural Lipids in Polymeric Membranes 

There are several methods to selectively open up closed polymeric membrane compartments in order to release entrapped substances (Fig. 37). For uncorking a polymerized vesicle, its membrane has to contain destabilizable areas which could possibly be opened up by variation of pH 70), temperature increase71), photochemical destabilization 72), or enzymatic processes. Such an enzymatic process is the hydrolysis of a natural phospholipid by phospholipase A2 (Fig. 38). This enzyme cleaves the ester bond in position two of a natural phosphoglyceride producing a lysophospholipid and a fatty acid which are both water soluble. This leads to complete destruction of the membrane. 

The action of phospholipase A2 on mixed monolayers of natural and polymerizable lipids can be measured under constant surface pressure by the contraction of the monolayer as a function of time as depicted schematically in Fig. 39. It turns out that the chief parameter influencing the enzymatic activity is the miscibility of the lipid components and not the fact whether the film is polymerized or not. In mixed and demixed membranes the enzyme is able to hydrolyze the natural lipid component, but with considerable differences in the hydrolizing rate (Fig. 40). A pure dilauroyllecithin (DLPC) monolayer is completely hydrolyzed in a few minutes after injecting the enzyme 

Preliminary experiments with 6-CF loaded mixed vesicles of dipalmitoylphosphat-idylcholine (DPPC) and diacetylenic lipid (22) exhibit only a very slow release of 6-CF after the addition of phopholipase A2. Reasons for this may be headgroup interactions which decrease the enzymatic activity or the fact that multilamellar vesicles were used for this study. Further experiments will be carried out with giant liposomes (visible under the light microscope) which are unilamellar and provide a higher enclosure percentage and low surface curvature (see 4.3.2). 

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In addition to enzymatic hydrolysis of natural lipids in polymeric membranes as discussed in chapter 4.2.2., other methods have been applied to trigger the release of vesicle-entrapped compounds as depicted in Fig. 37. Based on the investigations of phase-separated and only partially polymerized mixed liposomes 101, methods to uncork polymeric vesicles have been developed. One specific approach makes use of cleavable lipids such as the cystine derivative (63). From this fluorocarbon lipid mixed liposomes with the polymerizable dienoic acid-containing sulfolipid (58) were prepared in a molar ratio of 1 9 101115>. After polymerization of the matrix forming sulfolipids, stable spherically shaped vesicles are obtained as demonstrated in Fig. 54 by scanning electron microscopy 114>. 

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