Assessment of Fatty Acid-Specific Lipolysis by In Vitro Digestion and GC-FID

The nutritional relevance of food compositional data could be improved by taking the bioaccessibility of these constituents into account. A lack of routine methods to assess the bioaccessibility of fatty acids (FAs) in food is one of the limiting factors of doing so. An analytical protocol is proposed for routine assessment of the extent of lipolysis via in vitro digestion simulation methods in food products. The established method provides specific information on each FA individually. Steps of the protocol including the Bligh and Dyer chloroform/methanol/water extraction of esterified and free FAs from in vitro digesta, methyl ester derivatization, and GC-FID analysis were specifically tailored to help routine work and were harmonized with the Infogest in vitro digestion simulation protocol (both v1.0 and v2.0). The method was applied to assess the degree of FA-specific lipolysis in a baked fish (carp) meal and the results showed that the FA composition of the original food significantly differed from that of the distribution of FFAs in the digesta. The use of gastric lipase (in Infogest v2.0 protocol) increased total FA release by 9.5% and its specific impact on palmitic acid was the most prominent.

Dynamic In Vitro Gastric Digestion of Sheep Milk: Influence of Homogenization and Heat Treatment

Milk is commonly exposed to processing including homogenization and thermal treatment before consumption, and this processing could have an impact on its digestion behavior in the stomach. In this study, we investigated the in vitro gastric digestion behavior of differently processed sheep milks. The samples were raw, pasteurized (75 °C/15 s), homogenized (200/20 bar at 65 °C)–pasteurized, and homogenized–heated (95 °C/5 min) milks. The digestion was performed using a dynamic in vitro gastric digestion system, the human gastric simulator with simulated gastric fluid without gastric lipase. The pH, structure, and composition of the milks in the stomach and the emptied digesta, and the rate of protein hydrolysis were examined. Curds formed from homogenized and heated milk had much looser and more fragmented structures than those formed from unhomogenized milk; this accelerated the curd breakdown, protein digestion and promoted the release of protein, fat, and calcium from the curds into the digesta. Coalescence and flocculation of fat globules were observed during gastric digestion, and most of the fat globules were incorporated into the emptied protein/peptide particles in the homogenized milks. The study provides a better understanding of the gastric emptying and digestion of processed sheep milk under in vitro gastric conditions.

Gastric lipase can significantly increase lipolysis and carotenoid bioaccessibility from plant food matrices in the harmonized INFOGEST static in vitro digestion model

Gastrointestinal digestion of carotenoids has received much attention, as these lipophilic compounds have been related to several health benefits. Most commonly, static digestion models such as the consensus INFOGEST model...

Effects of recombinant human gastric lipase and pancreatin during in vitro pediatric gastro-intestinal digestion

The aim of this study was to implement a gastric digestion step using recombinant human gastric lipase (rHGL) in an in vitro pediatric gastro-intestinal digestion model to achieve a physiologically...

Crystal structure of human lysosomal acid lipase and its implications in cholesteryl ester storage disease

Lysosomal acid lipase (LAL) is a serine hydrolase that hydrolyzes cholesteryl ester (CE) and TGs delivered to the lysosomes into free cholesterol and fatty acids. LAL deficiency due to mutations in the LAL gene (LIPA) results in accumulation of TGs and cholesterol esters in various tissues of the body leading to pathological conditions such as Wolman’s disease and CE storage disease (CESD). Here, we present the first crystal structure of recombinant human LAL (HLAL) to 2.6 Å resolution in its closed form. The crystal structure was enabled by mutating three of the six potential glycosylation sites. The overall structure of HLAL closely resembles that of the evolutionarily related human gastric lipase (HGL). It consists of a core domain belonging to the classical α/β hydrolase-fold family with a classical catalytic triad (Ser-153, His-353, Asp-324), an oxyanion hole, and a “cap” domain, which regulates substrate entry to the catalytic site. Most significant structural differences between HLAL and HGL exist at the lid region. Deletion of the short helix, 238NLCFLLC244, at the lid region implied a possible role in regulating the highly hydrophobic substrate binding site from self-oligomerization during interfacial activation. We also performed molecular dynamic simulations of dog gastric lipase (lid-open form) and HLAL to gain insights and speculated a possible role of the human mutant, H274Y, leading to CESD.