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  • Competition of Thermomyces lanuginosus lipase with its hydrolysis products at the oil-water interface.

Competition of Thermomyces lanuginosus lipase with its hydrolysis products at the oil-water interface.

Colloids and surfaces. B, Biointerfaces (2016-10-23)
Marco Muth, Stefanie Rothkötter, Steven Paprosch, Reiner P Schmid, Klaus Schnitzlein
摘要

Lipase-catalyzed hydrolysis of triglycerides yields glycerol and free fatty-acids, provided that the enzyme is non-regioselective. For an Sn-1,3 regioselective enzyme, such as lipase from Thermomyces lanuginosus, the final product is no longer glycerol but Sn-2 monoglyceride instead. However, surface active molecules generated by lipolysis may have a detrimental effect on the interfacial biocatalysis since it is known that low molecular weight surfactants can displace proteins from interfaces. By using drop profile analysis tensiometry, we evaluated the interfacial properties of the lipase-generated molecules and their competitive effect on the adsorption behavior of the lipase and on the proceeding lipolysis. Our results show that even at concentration ratios of 8.64×10-4M (Sn-2 monoglyceride) to 2.5×10-7M (lipase), the final interfacial pressure values are very similar as for the system containing the lipase alone (i.e. ∼26 mN/m). This is a strong indication that monoglycerides, as the most interfacially active products generated during regioselective lipolysis, are expelled from the oil-water interface by the lipase. We attribute this effect to intermolecular lipase-lipase interactions, resulting in a low desorption probability of the lipase. For low oleic acid concentrations, the interfacial tension is solely determined by the lipase, while for higher concentrations, lipase and oleic acid both contribute to the tension values. We propose a hypothesis based on the preferential interaction of oleic acid molecules with hydrophobic sites on the lipase. The pH dependence of the adsorption rate and the interfacial activity of the lipase were also investigated.

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甘油三油酸酯, European Pharmacopoeia (EP) Reference Standard