No description available.

Yes / We report results of atomistic molecular dynamics simulations of an industrially-relevant, exemplar triacylglycerol (TAG), namely tristearin (TS), under aqueous conditions, at different temperatures and in the presence of an anionic surfactant, sodium dodecylbenzene sulphonate (SDBS). We predict the TS bilayers to be stable and in a gel phase at temperatures of 350 K and below. At 370 K the lipid bilayer was able to melt, but does not feature a stable liquid–crystalline phase bilayer at this elevated temperature. We also predict the structural characteristics of TS bilayers in the presence of SDBS molecules under aqueous conditions, where surfactant molecules are found to spontaneously insert into the TS bilayers. We model TS bilayers containing different amounts of SDBS, with the presence of SDBS imparting only a moderate effect on the structure of the system. Our study represents the first step in applying atomistic molecular dynamics simulations to the investigation of TAG-aqueous interfaces. Our results suggest that the CHARMM36 force-field appears suitable for the simulation of such systems, although the phase behaviour of the system may be shifted to lower temperatures than is the case for the actual system. Our findings provide a foundation for further simulation studies of the TS-aqueous interface. / veski

In the present work, we focused our attention on triacylglycerols. Their phase behavior strongly influences production processes of products based on fats and oils. However, the mechanisms controlling such behavior are not well understood. Hence, we decided to utilize computer simulation to gain more understanding in this matter. Using molecular dynamics simulation, we tried to mimic the thermal transition of triacylglycerols from liquid to solid state in order to shed more light on the crystallization of these species. The main conclusions are that due to the large time scale of the crystallization process, computer resources and time available for this work were not sufficient to simulate the full phase transition of triacylglycerols. Nevertheless, we managed to observe the first structural changes that drive the crystallization, i.e., the strong tendency of oxygen atoms to group and isolate themselves from the hydrocarbon chains. The successfully created molecular models are now ready for further investigations.

published_or_final_version / Chemistry / Master / Master of Philosophy

No description available.

Thesis (M. Phil.)--University of Hong Kong, 1996. / Includes bibliographical references (leaf 195-203).

Triacylglycerols are the predominant molecules of energy storage in eukaryotes. Triacylglycerol synthesis is catalyzed by acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymes, DGAT1 and DGAT2. Although the use of molecular tools, including targeted disruption of either DGAT enzyme, has shed light on their metabolic functions, little is known about the mechanisms responsible for regulating DGAT activity. Several lines of evidence from previous studies have suggested that DGAT1, but not DGAT2, is subject to regulation by phosphorylation and that protein kinase A (PKA)-dependent pathways are likely involved. In this study, the role of PKA in regulating DGAT activity and triacylglycerol synthesis during lipolysis was investigated. By using 3T3-L1 adipocytes, in vitro DGAT activity was shown to increase 2 fold during lipolysis. This data suggests that PKA might phosphorylate and activate DGAT1 during lipolysis to promote the recycling/re-esterification of excessive free fatty acids into triacylglycerols before they reach toxic levels within the cell. Additionally, high-performance liquid chromatography electrospray ionization mass spectrometry/mass spectrometry was exploited to identify PKA phosphorylation sites of DGAT1, and serine-17, -20 and -25 were identified as potential PKA phosphorylation sites using this methodology. The functional importance of these three potential phosphorylation sites was examined. Mutations of these sites to alanines (to prevent phosphorylation) or aspartates (to mimic phosphorylation) gave rise to enzymes functioning similarly to wild-type DGAT1. These phosphorylation sites appeared to be functionally silent as they were not involved in regulating DGAT1 activity, multimer formation, or enzyme stability. However, PKA phosphorylation at these three sites seemed to play a role in affinity of DGAT1 for its diacylglycerol substrate. These results indicate the existence of other unidentified, functionally active PKA phosphorylation sites or phosphorylation sites of other kinases, which are involved in regulating DGAT1.

Triacylglycerols are the predominant molecules of energy storage in eukaryotes. Triacylglycerol synthesis is catalyzed by acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymes, DGAT1 and DGAT2. Although the use of molecular tools, including targeted disruption of either DGAT enzyme, has shed light on their metabolic functions, little is known about the mechanisms responsible for regulating DGAT activity. Several lines of evidence from previous studies have suggested that DGAT1, but not DGAT2, is subject to regulation by phosphorylation and that protein kinase A (PKA)-dependent pathways are likely involved. In this study, the role of PKA in regulating DGAT activity and triacylglycerol synthesis during lipolysis was investigated. By using 3T3-L1 adipocytes, in vitro DGAT activity was shown to increase 2 fold during lipolysis. This data suggests that PKA might phosphorylate and activate DGAT1 during lipolysis to promote the recycling/re-esterification of excessive free fatty acids into triacylglycerols before they reach toxic levels within the cell. Additionally, high-performance liquid chromatography electrospray ionization mass spectrometry/mass spectrometry was exploited to identify PKA phosphorylation sites of DGAT1, and serine-17, -20 and -25 were identified as potential PKA phosphorylation sites using this methodology. The functional importance of these three potential phosphorylation sites was examined. Mutations of these sites to alanines (to prevent phosphorylation) or aspartates (to mimic phosphorylation) gave rise to enzymes functioning similarly to wild-type DGAT1. These phosphorylation sites appeared to be functionally silent as they were not involved in regulating DGAT1 activity, multimer formation, or enzyme stability. However, PKA phosphorylation at these three sites seemed to play a role in affinity of DGAT1 for its diacylglycerol substrate. These results indicate the existence of other unidentified, functionally active PKA phosphorylation sites or phosphorylation sites of other kinases, which are involved in regulating DGAT1.

<p> Edible oils consist primarily of triacylglycerols (or TAGs), which are triesters of glycerol and fatty acids. Determination of the TAG compositions of edible oils is becoming more important, given the economic value of these oil products and the increasing incidence of adulterating high quality oils with poorer quality oils. In this study we report the development of an analytical protocol using positive ion infusion electrospray mass spectrometry (ESI/MS) in conjunction with tandem mass spectrometry which affords both identification and quantification of TAGs in edible oils samples.</p> <p> This thesis reports a simple, comprehensive and quantitative method for the analysis of TAGs in edible oils in which the optimized method involves the infusion of an oil sample in chloroform:methanol (1:1) solution (~10-15 μg/mL of oil) in the presence of 0.5 mM LiCl. A sequence of corrections were applied to the raw peak area data of the TAG molecular ions, [M+Li]+, to account for: (1) normalization of peak area data using three internal standards, (2) peak area contributions of M+2 isotopic peaks of TAGs with one more degree of unsaturation and (3) peak area contributions of LiCl adduct ions, [M+Li+LiCl]+, when applicable. The major correction involved multiplication to a given TAG peak area by the appropriate electrospray relative response factor (RRF) for that TAG. The RRFs for all TAGs containing between 48 and 63 carbons in their fatty acyl chains and between 0 and 9 degrees of unsaturation were extrapolated from experimentally determined response factors of a series of standards. The RRFs were found to decrease by 6.7% for each additional acyl chain carbon but increased by 18.6% for each double bond. Comparison of these calculated RRFs to reported RRFs for a series of TAG standards showed an excellent correlation (1.06% ± 10.20% RSD).</p> <p> The use of Li+ in TAG analysis followed from the reports by Hsu and Turk [93] and Han and Gross [18] which showed that Li+ afforded more intense MS, and particularly MS/MS, spectra than either H+ or Na+. The enhanced intensities in MS/MS spectra (determined using a triple quadrupole mass spectrometer) were critical for the identification of TAGs, including the identity of the fatty acyl group located at the sn-2 position. However, this method cannot distinguish unambiguously between isobaric TAGs. This methodology was applied to the profiling of a number of edible oils including canola, olive, sesame, grape seed, walnut and hemp seed oils. The major TAGS in these samples contained 52, 54 and 56 carbons with between 0 and 11 degrees on unsaturation in a given TAG. There were minor amounts of TAGs containing 50, 55 and 57 carbons.</p> <p> The ability of this method to determine quantitatively the number of degrees of unsaturation in an oil sample was tested by examining a series of partially hydrogenated canola oil samples kindly provided by Bunge Canada. Five oil samples derived from a single feedstock with differing numbers of degrees of unsaturation, measured as iodine values, were subjected to our analytical method. The measured iodine values were compared to iodine values calculated from the number of degrees of unsaturation obtained by our MS-based method. The slope of this correlation was 1.10 with an R^2 = 0.995. Overall, this method is much simpler and more accurate than the protocol described by Han and Gross [18]. This methodology will be applied as routine method for the analysis of TAGs in biological samples such as blood samples.</p> / Thesis / Master of Science (MSc)

Fat bloom in chocolates is the gray-white discoloration and dullness that can occur on the surface of the confectionery. Fat bloom is a common quality defect that can result from temperature fluctuations during storage. Chocolates candies with peanuts or other nut fillings are more prone to fat bloom compared to plain chocolates, due to a release of incompatible nut oils into the chocolate matrix. The overall goal of this study was to determine if differences in triacylglycerol (TAG) composition and rheological properties of high, medium, and normal oleic peanuts influence fat bloom formation. All three peanut varieties showed high concentrations of triolein. Normal oleic peanuts had a slightly higher trilinolein than high and medium oleic peanuts, which contained trilinolein in trace amounts. Peanut pastes from the three peanut varieties all had a minimum apparent yield stress, and all pastes showed varying degrees of shear thinning. The apparent yield stress of high and normal oleic pastes was higher than the apparent yield stress of medium oleic paste. The absolute value of the flow index behavior was 1 for the high oleic peanut paste, suggesting friction in the experimental apparatus, even with use of Teflon plates. The peanut chocolate candies took around 45 days for significant dulling of the chocolates with temperature cycling between 26-29 °C approximately every 26 hours. Optical microscopy scans showed differences in glossiness and surface textural attributes of the unbloomed and bloomed peanut chocolate confectionery. Consumer evaluation showed some differences in the glossiness and significant differences in surface texture of unbloomed and bloomed chocolates. A majority (62%) of the survey respondents had seen whitish discoloration in chocolates and 40% of the respondents thought this is because the chocolate had grown old. / Ph. D.