Aiqian Ye

Structuring food emulsions in the gastrointestinal tract to modify lipid digestion

The importance of nutrient lipids in the human diet has led to major advances in understanding the mechanisms of lipid digestion and absorption. With these advances has come new recognition that the matrix in which lipids are presented (i.e. food structure) in the diet could influence the rate of lipid digestion and hence the bioavailability of fatty acids. As a consequence, there is growing interest in understanding how food material properties can be manipulated under physiological conditions to control the uptake of lipids and lipid-soluble components. The lipids in many, if not most, processed foods are normally present as emulsions, which can be end products in themselves or part of a more complex food system. In this review, we discuss the formation and properties of oil-in-water (O/W) emulsions, especially how these emulsions are modified as they traverse through the gastrointestinal tract. Among other factors, the changes in the nature of the droplet adsorbed layer and the droplet size play a major role in controlling the action of lipases and lipid digestion. Greater knowledge and understanding of how the digestive system treats, transports and utilizes lipids will allow the microstructural design of foods to achieve a specific, controlled physiological response.

Characterization of protein components of natural and heat-treated milk fat globule membranes

The proteins associated with the milk fat globule membrane (MFGM), isolated from early, mid and late season whole milks, were characterized using one- and two-dimensional SDS-PAGE under reducing and non-reducing conditions. In some experiments, MFGM separated from fresh whole milk was suspended in simulated milk ultrafiltrate (SMUF) and heated at various temperatures and times. SDS-PAGE under reducing conditions followed by staining with Coomassie blue showed the presence of about 37 protein bands, ranging in molecular weight from 15 to 200 kDa. SDS-PAGE under non-reducing conditions showed only about 25 distinct bands and the intensity of xanthine oxidase and butyrophilin bands was much less, while the intensity of PAS 6/7 band was similar compared with the reduced SDS-PAGE. Two-dimensional SDS-PAGE showed that the protein complexes that remained at the top of non-reducing gel were resolved into mostly xanthine oxidase and butyrophilin with a small proportion of PAS 6. These results indicate that xanthine oxidase and butyrophilin may be complexed via intermolecular disulfide bonds in the natural MFGM, although it is not possible to differentiate the individual protein distributions within these aggregates. It was found that the total protein content (mg/g fat) of MFGM and the percentage of xanthine oxidase and butyrophilin in early and late season MFGM were higher than that of mid season MFGM. In heated samples (above 60°C), xanthine oxidase and butyrophilin interacted further to form higher molecular weight protein complexes, while PAS 6/7 was relatively heat stable.

Interfacial composition and stability of emulsions made with mixtures of commercial sodium caseinate and whey protein concentrate.

The interfacial composition and the stability of oil-in-water emulsion droplets (30% soya oil, pH 7.0) made with mixtures of sodium caseinate and whey protein concentrate (WPC) (1:1 by protein weight) at various total protein concentrations were examined. The average volume-surface diameter (d32) and the total surface protein concentration of emulsion droplets were similar to those of emulsions made with both sodium caseinate alone and WPC alone. Whey proteins were adsorbed in preference to caseins at low protein concentrations (<3%), whereas caseins were adsorbed in preference to whey proteins at high protein concentrations. The creaming stability of the emulsions decreased markedly as the total protein concentration of the system was increased above 2% (sodium caseinate >1%). This was attributed to depletion flocculation caused by the sodium caseinate in these emulsions. Whey proteins did not retard this instability in the emulsions made with mixtures of sodium caseinate and WPC.

In Vitro Digestion of β-Lactoglobulin Fibrils Formed by Heat Treatment at Low pH

Extensive studies have been done on beta-lactoglobulin (beta-Lg) fibrils in the past decade due to their potential as functional food ingredients, gelling agents, and encapsulation devices etc. (van der Goot, A. J.; Peighambardoust, S. H.; Akkermans, C.; van Oosten-Manski, J. M. Creating novel structures in food materials: The role of well-defined shear flow. Food Biophys. 2008, 3(2), 120-125 and Loveday, S. M.; Rao, M. A.; Creamer, L. K.; Singh, H. Factors affecting rheological characteristics of fibril gels: The case of beta-lactoglobulin and alpha-lactalbumin. J. Food Sci. 2009, 74 (3), R47-R55). However, most of the studies focus on the formation and mechanism of the fibrils. Little is known about fibril digestibility to date. In this work, in vitro pepsin digestion of bovine beta-lactoglobulin (beta-Lg) fibrils in simulated gastric fluid was investigated using thioflavin T fluorescence photometry, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, size-exclusion chromatography, matrix-assisted laser desorption/ionization mass spectrometry, and transmission electron microscopy (TEM). The fibrils were formed by heating beta-Lg solutions at 80 degrees C and pH 2.0 for 20 h. The fibrils were found to be digested completely by pepsin within 2 min, when long, straight fibrils were no longer observed by TEM. The peptides in the fibrils (2000-8000 Da) could be digested to smaller peptides (mostly <2000 Da) by pepsin. The peptides in the fibrils were believed to be more susceptible for pepsin to access and attack because of their hydrophobic nature. For comparison purposes, solutions of beta-Lg heated at neutral pH (pH 7.4) were also studied under the same conditions.

Structural changes of bovine milk fat globules during in vitro digestion

An in vitro digestion model that simulated gastric and intestinal fasting conditions was used to monitor the physical, chemical, and structural changes of fat globules from raw bovine milk. During in vitro gastric digestion, the fat globules were stable under low-acidic conditions. Some peptides and β-lactoglobulin were resistant to proteolysis by pepsin. Phospholipids, proteins, and peptides stabilized the globules in the stomach model. During in vitro intestinal digestion, most of the β-lactoglobulin and residual peptides were hydrolyzed by trypsin and chymotrypsin, and the lipolytic products, released from the hydrolysis of the triglyceride core of the globules, led to destabilization and coalescence of the globules. By accumulating at the surface of the fat globules, the lipolytic products formed a lamellar phase and their solubilization by bile salts resulted in the formation of disk-shaped micelles. This study brings new interesting insights on the digestion of bovine milk.

In vitro gastric digestion of heat-induced aggregates of β-lactoglobulin.

In vitro gastric digestion of heat-induced aggregates of β-lactoglobulin (β-LG) in simulated gastric fluid was investigated using sodium dodecyl sulfate-PAGE (under nonreducing and reducing conditions), native PAGE, 2-dimensional electrophoresis, and size exclusion chromatography. Heating at 90°C significantly increased the digestibility of β-LG, with a high initial digestion rate followed by a relatively constant rate of digestion at a high enzyme:substrate (E:S) ratio of 3:1. At a low E:S ratio (1:6), the rate of digestion of β-LG was slower, and intermediate- and low-molecular-weight species could be seen. The high-molecular-weight nonnative aggregates (e.g., pentamers, tetramers, and trimers) were digested relatively rapidly, whereas some of the nonnative dimers were resistant to digestion and others were digested rapidly. The intermediate-molecular-weight species (21 to 23 kDa) were digested slowly. The digestibility of nonnative β-LG aggregates varied significantly depending on the E:S ratio and the types of aggregate. Further investigation is necessary to identify and characterize slowly digested dimers and species of intermediate molecular weight.

Proteolysis of milk fat globule membrane proteins during in vitro gastric digestion of milk.

The influence of gastric proteolysis on the physicochemical characteristics of milk fat globules and the proteins of the milk fat globule membrane (MFGM) in raw milk and cream was examined in vitro in simulated gastric fluid (SGF) containing various pepsin concentrations at pH 1.6 for up to 2h. Apparent flocculation of the milk fat globules occurred in raw milk samples incubated in SGF containing pepsin, but no coalescence was observed in either raw milk samples or cream samples. The changes in the particle size of the fat globules as a result of the flocculation were dependent on the pepsin concentration. Correspondingly, the physical characteristics of the fat globules and the composition of the MFGM proteins in raw milk changed during incubation in SGF containing pepsin. The major MFGM proteins were hydrolyzed at different rates by the pepsin in the SGF; butyrophilin was more resistant than xanthine oxidase, PAS 6, or PAS 7. Peptides with various molecular weights, which altered with the time of incubation and the pepsin concentration, were present at the surfaces of the fat globules.

In vivo digestion of bovine milk fat globules: Effect of processing and interfacial structural changes. I. Gastric digestion

The aim was to study the in vivo gastric digestion of fat globules in bovine cream from raw, pasteurised or pasteurised and homogenised milk. Fasted rats were gavaged once and chyme samples were collected after 30, 120 and 180 min post-gavage. Proteins from raw (RC) and pasteurised (PC) creams appeared to be digested faster and to a greater extent. Free fatty acids (FAs) increased throughout the 3h postprandial period. Short and medium chain FAs were released more rapidly than long chain FAs which were hydrolysed to a greater degree from PC. The size of the fat globules of all creams increased in the stomach. Protein aggregates were observed in pasteurised and homogenised cream chyme. Protrusions, probably caused by the accumulation of insoluble lipolytic products, appeared at the surface of the globules in RC and PC chyme. Overall, PC proteins and lipids appeared to be digested to a greater extent.

Influence of gastric digestive reaction on subsequent in vitro intestinal digestion of sodium caseinate-stabilized emulsions

In this study, in vitro intestinal lipid digestion and the physicochemical and microstructural changes of sodium caseinate-stabilized emulsions were examined after the emulsions had been digested in a model simulated gastric fluid containing pepsin for different times. The average size, size distribution, microstructure, proteolysis of interfacial proteins and lipolysis of the emulsion droplets were monitored as a function of digestion time. The emulsion droplets underwent extensive droplet flocculation, with some coalescence together with proteolysis of interfacial proteins, in simulated gastric fluid, resulting in changes in the droplet size and the microstructure of the emulsions. In general, digestion in simulated gastric fluid containing pepsin accelerated coalescence of the emulsion droplets during subsequent digestion in simulated intestinal fluid containing pancreatic lipase. However, the changes in the size, the microstructure and the proteolysis of the interfacial proteins of the emulsions under gastric conditions did not influence the rate and the extent of lipid digestion in the subsequent intestinal environment.

Stability during in vitro digestion of lactoferrin-loaded liposomes prepared from milk fat globule membrane-derived phospholipids

Liposomes loaded with positively charged lactoferrin (LF) were prepared from milk fat globule membrane-derived phospholipids using a thin-layer dispersion method. The entrapment efficiency of LF in the liposomes and the stability during in vitro gastrointestinal digestion were characterized and examined using dynamic light scattering, transmission electron microscopy, and PAGE. The entrapment efficiency of LF encapsulated in the liposomes was about 46%. The entrapped LF remained unchanged as a function of time and pepsin concentration when the liposome samples were digested in a simulated gastric environment, suggesting that the liposomes prepared from milk fat globule membrane-derived phospholipids were stable and protected the entrapped LF from pepsin hydrolysis. In simulated intestinal fluid, the entrapped LF was more susceptible to hydrolysis by the protease in pancreatin, as shown by changes in the diameter and membrane structure of the liposomes. The release of free fatty acids from the liposomes during digestion in simulated intestinal fluid revealed that the phospholipids in the liposomes were partly hydrolyzed by pancreatic lipase. It was suggested that liposomes may prevent the gastric degradation of LF and reduce the rate of hydrolysis of LF in intestinal conditions.