Cutinase binding and activity at the triolein–water interface monitored by oil drop tensiometry

Flipsen, J.A.C.; Krämer, Reinhard; Duijnhoven, John P. M. van; Hijden, H.T.W.M. van der; Egmond, Maarten R.; Verheij, Hubertus M.

doi:10.1016/s0009-3084(98)00072-3

Cited by 9 publications

(4 citation statements)

References 27 publications

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“…Injection of colipase and lipase did not show any immediate change in interfacial tension, but demonstrated a lag time before the interfacial tension started to decrease until the oil drop detached from the needle (Figure B). The decrease in interfacial tension was a combined effect of enzyme adsorption and lipolysis product formation at the interface. , The maximum rate of the decrease was determined from the maximum slope of the linear portion of the interfacial tension kinetic profile. The lag time was the duration from the addition of colipase and lipase to the time when the interfacial tension decreased linearly at the highest rate which was determined at the intercept between the maximum slope and the tangent of the interfacial tension plateau to the time-axis of the curve.…”

Section: Resultsmentioning

confidence: 99%

Modulating Pancreatic Lipase Activity with Galactolipids: Effects of Emulsion Interfacial Composition

et al. 2009

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It is widely known that the interfacial quality of lipid emulsion droplets influences the rate and extent of lipolysis. The aim of this work was to investigate the effect of two galactolipids, monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), adsorbed at the interface on in vitro digestibility of olive oil by porcine pancreatic lipase. The experiments were performed under simulated duodenal conditions in the presence of phosphatidylcholine (lecithin) and bile salts. It was found that emulsions prepared with DGDG had a longer lag phase prior to lipase activation with a decrease in lipolysis rate. In contrast, no inhibitory effect on lipase kinetics was observed in emulsions prepared with MGDG. We postulated that the larger headgroup and more tightly packed molecular organization of DGDG at the interface gave rise to steric hindrance that retarded colipase and lipase adsorption onto the substrate surfaces and hence delayed and reduced lipolysis. It was noted that the lag phase and lipolysis rate strongly depended on the DGDG/lecithin molar ratio in the systems: the higher the molar ratio, the longer the lag phase followed by a reduced lipolysis rate. The ability of DGDG to inhibit bile salt adsorption/displacement was also investigated. The results showed that bile salts did not completely displace DGDG from the interface, explaining the reason why DGDG still possessed inhibitory activity even in the presence of bile salts at a physiological relevant concentration. The results provide interesting insights into the influence of the galactolipid headgroup and lecithin on the emulsion interfacial quality which in turn regulates the lipolysis. The findings potentially could lead to the production of generic foods and drugs designed for regulating dietary fat absorption in the prevention and treatment of obesity and related disorders.

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Section: Resultsmentioning

confidence: 99%

Modulating Pancreatic Lipase Activity with Galactolipids: Effects of Emulsion Interfacial Composition

et al. 2009

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“…As compared with the other interfacial techniques, the oil‐drop tensiometer presents the unique advantage of being able to monitor lipase activities on natural long‐chain triacylglycerols at a closely controlled oil/water interface [63, 64]. However, the oil‐drop methodology requires the oil to be freed carefully from any natural tensioactive compounds such as free fatty acids and di‐ and mono‐acylglycerols because of the amphipathic character of these contaminants, which might decrease the initial interfacial tension.…”

Section: Lipase Assay Proceduresmentioning

confidence: 99%

Lipase assays for conventional and molecular screening: an overview

Gupta

Rathi

Gupta

et al. 2003

Biotech and App Biochem

160

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Lipases are versatile biocatalysts that can perform innumerable different reactions. Their enantio-, chemo- and stereo-selective nature makes them an important tool in the area of organic synthesis. Unlike other hydrolases that work in aqueous phase, lipases are unique as they act at the oil/water interface. Besides being lipolytic, lipases also possess esterolytic activity and thus have a wide substrate range. Hence, the lipase assay protocols hold a significant position in the field of lipase research. Lipase activity can be estimated using a wide range of assay protocols that differ in terms of their basic principle, substrate selectivity, sensitivity and applicability. As the value of these enzymes continues to grow and new markets are exploited, development of new or improved enzymes will be a key element in the emerging realm of biotechnology. Hence, development of faster and simpler protocols incorporating newer and more specific substrates is the need of the hour. In this endeavour, methods that could be adopted for molecular screening occupy an important position. Here, an overview of the lipase assay protocols is presented with emphasis on the assays that can be adopted for the molecular screening of these biocatalysts.

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“…Furthermore, the surface behaviour (interfacial binding) of lipases and mutants at the oil-water interface can be studied under similar conditions [166][167][168]. The oil-drop methodology requires the oil to be carefully freed from any natural tensioactive compounds such as FFAs and di-and monoglycerides because of the amphipathic character of these contaminants, which might decrease the initial interfacial tension.…”

Section: The Oil-drop Methodsmentioning

confidence: 99%