The milk fat globule membrane (MFGM) has undergone a renaissance in nutritional sciences. Recent understanding of how heterogenous, lateral milk polar lipids assemblies, enriched with sphingomyelin and cholesterol, form a liquid-ordered (L-o) phase, call assumptions about milk fat digestion into question. Clinical studies on the anticholesteremic effect of milk polar lipids, fundamental research on the structure and properties of L-o domains, and hypotheses on the sphingomyelin and cholesterol interactions that lead to the establishment of L-o structures are critically reviewed. Whereas previous reviews delivered overviews on such topics, this review critically considers the interplay of structure, digestion, and composition.

Sphingomyelin (SM) and cholesterol complex to form functional liquid-ordered (Lo) domains. It has been suggested that the detergent resistance of these domains plays a key role during gastrointestinal digestion of the milk fat globule membrane (MFGM), which is rich in both SM and cholesterol. Small-angle X-ray scattering was employed to determine the structural alterations that occur when milk sphingomyelin (MSM)/cholesterol, egg sphingomyelin (ESM)/cholesterol, soy phosphatidylcholine (SPC)/cholesterol, and milk fat globule membrane (MFGM) phospholipid/cholesterol model bilayer systems were incubated with bovine bile under physiological conditions. The persistence of diffraction peaks was indicative of multilamellar vesicles of MSM with cholesterol concentrations > 20 % mol, and also for ESM with or without cholesterol. The complexation of ESM with cholesterol is therefore capable of inhibiting the resulting vesicles from disruption by bile at lower cholesterol concentrations than MSM/cholesterol. After subtraction of background scattering by large aggregates in the bile, a Guinier fitting was used to determine changes in the radii of gyration (Rgs) over time for the biliary mixed micelles after mixing the vesicle dispersions with bile. Swelling of the micelles by phospholipid solubilization from vesicles was a function of cholesterol concentration, with less swelling of the micelles occurring as the cholesterol concentration was increased. With 40% mol cholesterol, the Rgs of the bile micelles mixed with MSM/cholesterol, ESM/cholesterol, and MFGM phospholipid/cholesterol were equal to the control (PIPES buffer + bovine bile), indicating negligible swelling of the biliary mixed micelles.

Olive cake, the solid byproduct of three-phase centrifugation olive oil production, has a high organic and polyphenol content, rendering it an environmental threat when landfilled as well as limiting its animal feed potential. This residue can be a good candidate for biomethane production due to its rich polysaccharide content (pectin, hemicellulose, and cellulose). Two strategies were compared to maximize biomethane production: destoning (i.e., removal of the seed fragments via mechanical means) and enzymatic pretreatment of the pulp. After 30 days of batch anaerobic digestion at 35 °C, both enzymatically pretreated and destoned olive cakes produced similar amounts of methane (~295 mL CH4/g volatile solids (VS)), 42% more than the control. A comparison of olive cake’s biomethane yields with a broad range of agricultural residues in the literature demonstrated its suitability for biomethane production. Additionally, the digestate recovered from the anaerobic digestion of olive cake had high Kjeldahl nitrogen contents (3.6%, db) and low polyphenol concentrations (0.02 mg gallic acid equivalent (GAE)/g), qualifying it as an ingredient for soil amendment. This study demonstrated olive cake can be diverted from landfills for second-generation biofuel production, and that the resulting digestate may have value for soil amendment.

Sphingomyelin (SM) and cholesterol complexation gives rise to detergent-resistant liquid-ordered domains. The persistence of these domains and subsequent mixed micelle formation was examined in the presence of bile under physiological digestive in vitro conditions for vesicles comprising either SM/cholesterol, porcine brain phosphatidylcholine (BPC)/cholesterol, or soy phosphatidylcholine (SPC)/cholesterol bilayers, the latter two systems having no liquid-ordered domains. Micellization of these digested phospholipid multilamellar vesicle systems was confirmed by transmission electron microscopy. Bovine bile was found to consist of large multilamellar sheets which subsumed phospholipid vesicles to form aggregated superstructures. Budding off from these superstructures were vesicle-to-micelle transition intermediates: unilamellar vesicles and cylindrical micelles. The presence of cholesterol (60/40 phospholipid/cholesterol mol/mol) delayed the initial rapid onset of digestion, but not for BPC and SPC vesicle systems. Acyl chain order/disorder before and after vesicle-to-micelle transition of all three phospholipid/cholesterol systems was examined using Raman spectroscopy. The addition of bovine bile to both PC/cholesterol vesicle systems reduced the overall ratio of acyl chain disorder to order. In SM/cholesterol vesicles with ≤ 20% mol cholesterol, only the lateral inter-acyl chain packing was reduced, whereas for SM/cholesterol vesicles with ≥ 30% mol cholesterol, a higher proportion of gauche-to-trans isomerization was apparent, demonstrating that SM/cholesterol complexes modify the acyl chain structure of micelles.

More than 16,000 tons of olive cake was produced in the United States in 2017. Olive cake is a by-product of olive oil extraction, which has limited animal feed potential, and poses an environmental threat when landfilled due to its high organic load and polyphenol content. This residue has potential for biofuel (bioethanol and biomethane) production because it is rich in polysaccharides such as pectin, hemicellulose, and cellulose. Yet, olive cake contains olive stones that can impede its conversion to biofuel. Therefore, two methods of destoning, centrifugation and screening by horizontal screw press, were first compared. Both methods removed an equal percentage of stones (95%), but centrifugation partitioned the majority (57 – 79%) of digestible solids (olive pulp) with the stones. Then, two strategies were compared to maximize both biomethane and bioethanol production; enzymatic conversion of insoluble to soluble carbohydrates and destoning by screening. After 30 days of anaerobic digestion at 35 °C, both the enzymatically pretreated and the destoned olive cakes produced similar amounts of methane (~295 mL CH4/g VS), 42% more than the control (209.5 mL CH4/g VS). The biogas produced was composed of 60-70% methane. A comparison of biomethane yields with a broad range of agricultural residues demonstrated olive cake’s suitability for biomethane production. The digestate, residue from the anaerobic digestion, have high Kjeldahl nitrogen content (3.6%, db) and low polyphenol concentration (0.02 mg GAE/g), which then qualify it as an ingredient for soil amendment. Ethanol production investigations showed that after 3 days of fermentation at 32 °C, only the destoned and enzymatically pretreated olive cake produced ethanol (1.3 mg/mL). Acetic acid, an inhibitor of ethanol production, was present in all samples broth, suggesting microbial contamination was present.