Lipase-catalyzed transesterification of phosphatidylcholine at controlled water activity

The incorporation of a free fatty acid into thesn-1 position of phosphatidylcholine by lipase-catalyzed transesterification was investigated. The thermodynamic water activity of both the enzyme preparation and the substrate solution was adjusted to the same value prior to the reaction. The reaction rate increased with increasing water activity but the yield of modified phosphatidylcholine decreased due to hydrolysis. By using a large excess of the free fatty acid (heptadecanoic acid), the hydrolysis reaction was slowed down, so a higher yield was obtained at a given degree of incorporation. The best results were obtained withRhizopus arrhizus lipase immobilized by adsorption on a polypropylene support. With this preparation, a yield of 60% and nearly 50% incorporation of heptadecanoic acid (100% incorporation in thesn-1 position) was obtained at a water activity of 0.064. The enzyme preparation had good operational stability and position specificity. Little incorporation (<1%) was observed in thesn-2 position, when almost all the fatty acid in thesn-1 position was exchanged.     

Associations between skeletal muscle properties, physical fitness, physical activity and coronary heart disease risk factors in men

High physical fitness and physical activity are associated with favourable lipid levels, especially a high level of high density lipoprotein cholesterol (HDL-C). A person's skeletal muscle properties, metabolism and percentage of different muscle fibres (ST-%), which may modify coronary heart disease (CHD) risk factors, such as serum insulin, obesity and serum sex hormones may also influence his fitness level and leisure-time physical activity. We studied the associations of physical fitness, physical activity and ST-% with serum lipids and lipoproteins in 72 healthy men. Their parameters were compared with those of 20 men with defined CHD. Significant interrelationships between ST-%, fitness and leisure-time physical activity index (LTPAI) were observed. Multiple regression analysis showed that ST-%, fitness and leisure-time physical activity explained about 32% of the variation in HDL-C in the healthy men. In healthy men ST-% correlated positively with fitness (r(s) = 0.62, P < 0.001) and with LTPAI (r(s) = 0.62, P < 0.001). Fitness level also correlated significantly with LTPAI (r(s) = 0.81, P < 0.001). Serum insulin showed negative associations with ST-% (r(s) = -0.63, P < 0.001) and fitness (r(s) = -0.54, P < 0.001) and LTPAI (r(s) = -0.62, P < 0.001). Free fraction of testosterone correlated negatively with serum HDL-C level (r(s) = -0.34, P < 0.01), with fitness (r(s) = -0.41, P < 0.001) and with LTPAI (r(s) = -0.54, P < 0.001). In sedentary men with the lowest fitness and physical activity the mean of ST-% (45%) was similar to that in CHD patients (44%). However, ST-% in men in the highest tertile of physical activity and fitness (68%) was significantly higher than in CHD patients and in men in the lowest tertile of physical activity and fitness. Skeletal muscle enzyme activity in lipid metabolism was significantly lower in both CHD patients and in sedentary and low-fit men than that in fitter and physically active men. The present data imply that skeletal muscle properties are important determinants of risk profiles, such as physical activity, fitness and serum lipid and lipoprotein patterns. Although fitness is a graded, independent predictor of mortality from CHD, a relatively high fitness level is not enough. This was clearly observed in the clustering analysis, in which the healthy men, according to their ST-%, fitness, leisure-time physical activity and serum sex hormone binding globulin (SHBG), fell into three natural groups: (i) Inactive men with lowest ST-% (mean 42%), lowest fitness (10.7 METs) and lowest HDL-C (1.36 mm/l); (ii) Fit men with high ST-% (66%), high fitness (14.5 METs) and moderately high HDL-C (1.54 mol/l); (iii) Active men with high ST-% (66%), highest fitness (14.9 METs) and highest serum HDL (1.83 mmol/l). The results support the idea that both fitness and physical activity give further protection against CHD by modifying risk factors. Our findings also suggest that skeletal muscle properties should be considered in the studies which assess CHD risk factors and their modifications especially in the field of health-related fitness.     

Processing of rye bran influences both the fermentation of dietary fibre and the bioconversion of lignans by human faecal flora in vitro

Plant lignans are converted to mammalian forms, enterodiol and enterolactone, in the colon. Enhanced plasma or urinary enterolactone levels have been associated with lowered risk of cardiovascular diseases and breast cancer. Processed rye bran and its fractions were compared to ascertain the fermentation rates of fermentable carbohydrates and the bioconversion of lignans. Rye bran was extruded and treated with a food‐grade xylanase. Part of the xylanase‐treated rye bran was separated into a soluble rye bran extract and an insoluble residue, and a part of the xylanase‐treated rye bran was freeze‐dried without separation. All the samples were digested by pepsin and pancreatin and subsequently fermented with a human faecal inoculum in vitro. The consumption of carbohydrates, the productions of short‐chain fatty acids (SCFA), enterodiol and enterolactone were followed as a function of time. The soluble rye bran extract showed the fastest fermentation rate and the highest extent of fermentation determined as the consumption of neutral sugar residues (arabinose, xylose and glucose), the production of SCFA and the formation of enterodiol and enterolactone. Xylanase treatment enhanced the fermentation rate of extruded rye bran. An even a higher fermentation rate was observed for rye bran extract containing soluble carbohydrates. The amount of enterolactone precursors in rye seemed to be too low for enterolactone formation using an amount of substrate suitable for carbohydrate fermentation. However, xylanase treatment enhanced the availability of plant lignans from rye bran, as enterodiol formation was increased by the use of xylanase. Copyright © 2005 Society of Chemical Industry     

Polar Lipids Reduce In Vitro Duodenal Lipolysis Rate of Oat Oil and Liquid Oat Base Products

Alternative ways for increased appetite control are today widely sought for due to the growing global health issues connected to obesity. In in vivo studies, oat has been proven an attractive candidate for inducing satiety. Oat is rich in polar lipids, of which the galactolipids are especially interesting, and a hypothesis is that these lipids play an important role for the ileal brake mechanism. In this study, the aim is to investigate the role of polar oat lipids on pancreatic lipolysis rate, using a pH-stat based in vitro digestion model of the duodenum. Lipolysis of oat oil, a mix of oat oil/rapeseed oil (RSO), as well as a liquid oat base (OB) simulating an oat drink with different polar lipid content are investigated, and compared with RSO as control. Increasing the polar lipid content of the product digested leads to a significantly decreased lipolysis rate, and this effect is even observed when mixing RSO with a low amount of oat oil (10%). The results support the hypothesis that polar lipids can delay lipolysis also in a complex, natural system like the liquid OB, and even a minor amount of oat lipids can have large effect on lipolysis rates. Practical applications: The number of studies connecting galactolipids with a decreasing effect on duodenal lipolysis is growing; however, the mechanism behind this phenomenon is still not clarified. Here, the same effect is seen in a complex, natural food system. These findings open up for interesting future food products, where inclusion of oat oil, even at low concentrations, can have a prolonging effect on satiety. Oat for human consumption is an increasing market, thanks to the positive health benefits oat has been connected to, in combination with the current trend toward climate-friendly plant-based options for meat and dairy products. It is believed that oat oil can be attractive as an ingredient in various food products, for example, protein bars and spreads. More studies are needed to confirm the results in vivo. However, a great potential is seen for the use of oat oil to enhance appetite control.     

Chemo‐enzymatic epoxidation of linoleic acid: Parameters influencing the reaction

The effect of reaction parameters on lipase‐mediated chemo‐enzymatic epoxidation of linoleic acid was investigated. Hydrogen peroxide was found to have the most significant effect on the reaction rate and degree of epoxidation. Excess of hydrogen peroxide with respect to the amount of double bonds was necessary in order to yield total conversion within a short time period, as well as at temperatures above 50 °C to compensate for hydrogen peroxide decomposition. However, prolonged incubation with high excess of hydrogen peroxide leads to the accumulation of peracids in the final product. The reaction rate increased also with increasing hydrogen peroxide concentration (between 10 and 50 wt‐%); however, at the expense of enzyme inactivation. Linoleic acid was completely epoxidized when used at a concentration of 0.5–2 M in toluene at 30 °C, while in a solvent‐free medium, the reaction was not complete due to the formation of a solid or a highly viscous oily phase, creating mass transfer limitations. Increasing the temperature up to 60 °C also improved the rate of epoxide formation.     

Partial purification and characterization of acylester hydrolase from Lupinus mutabilis

An alkaline acylester hydrolase was partially purified from germinated seeds of Lupinus mutabilis. Hydrolytic activity was absent in the crude extract of ungerminated lupine seed, but it increased and peaked at the fourth day in the germinating seedling. The purification scheme involved homogenization, centrifugation, acetone precipitation, anion exchange chromatography, pH precipitation, and hydrophobic interaction chromatography. The acylester hydrolase was purified 126-fold, and the overall activity yield was 10%. The molecular weight estimated by sodium dodecylsulfate-polyacrylamide gel electrophoresis was 60 kDa. The enzyme had an isoelectric point of 6.2 and showed maximal activity at pH 8.0. The enzyme showed good stability between pH 5.0 and 9.0. In the pH range 7.0–7.5, enzyme precipitation was observed. The enzyme was stable from 0 to 25°C for 5 h and at 45°C lost 50% of its activity in the same period of time. At higher temperatures, the enzyme showed low thermal stability. However, the highest initial activity was found to be at 45°C. Nonionic surfactants and cholic acid enhanced the activity of the enzyme. The activity was reduced by the addition of toluene and isooctane and increased by the addition of diethyl ether, acetonitrile, methanol, and pyridine. The activity was reduced by 37% in the presence of 1 mM Cu²⁺ ions. The enzyme-hydrolyzed triolein showing no positional specificity.     

Flavonoids, dietary-derived inhibitors of cell proliferation and in vitro angiogenesis

Consumption of a plant-based diet can prevent the development and progression of chronic diseases associated with extensive neovascularization, including solid malignant tumors. In previous studies, we have shown that the plant-derived isoflavonoid genistein is a potent inhibitor of cell proliferation and in vitro angiogenesis. In the present study, we report that certain structurally related flavonoids are more potent inhibitors than genistein. Indeed, 3-hydroxyflavone, 3',4'-dihydroxyflavone, 2',3'-dihydroxyflavone, fisetin, apigenin, and luteolin inhibited the proliferation of normal and tumor cells, as well as in vitro angiogenesis, at half-maximal concentrations in the low micromolar range. We have previously demonstrated that genistein concentrations in the urine of subjects consuming a plant-based diet is 30-fold higher than in subjects consuming a traditional Western diet. The wider distribution and the more abundant presence of flavonoids in the plant kingdom, together with the present results, suggest that flavonoids may contribute to the preventive effect of a plant-based diet on chronic diseases, including solid tumors.     

Intake of rye bread ileostomists increases ileal excretion of fiber polysaccharide components and organic acids but does not increase plasma or urine lignans and isoflavonoids

The excretion of starch, enzyme-resistant starch, dietary fiber components and organic acids (short-chain fatty acids plus lactic acid) as well as plasma and urine lignans and isoflavonoids was studied in eight ileostomists consuming mixed diets with wheat bread (low fiber diet) or rye bread (high fiber diet) in a crossover design. Average ileal excretions of enzyme-available starch were 3.5 g/d during the low fiber period and 4.1 g/d during the high fiber period. The excretion of enzyme-resistant starch was approximately the same (2.3 g/d) in both periods. In comparison with intake, similar amounts of total fiber residues were excreted both by subjects receiving the low fiber diet (3.4 g/d) and by those receiving the high fiber diet (2.7 g/d). However, subjects excreted significantly more of certain polysaccharide residues (fucose, galactose, and uronic acids) than they ingested. On average, the excretion of organic acids was 18.6 mmol/d during the low fiber period and 30.2 mmol/d during the high fiber period. No significant differences in plasma lignans were observed between the high fiber and the low fiber dietary periods. The present findings indicate that enzyme-available starch is highly digested and that a microbial breakdown of dietary fibers and probably other carbohydrates occurs in the small intestine. However, the bacterial activity in the ileostomists was not sufficient to cause an increased level in plasma lignans even when subjects consumed the high fiber rye diet.