Diatoms synthesize sterols by inclusion of animal and fungal genes in the plant pathway

Diatoms are ubiquitous microalgae that have developed remarkable metabolic plasticity and gene diversification. Here we report the first elucidation of the complete biosynthesis of sterols in the lineage. The study has been carried out on the bloom-forming species Skeletonema marinoi and Cyclotella cryptica that synthesise an ensemble of sterols with chemotypes of animals (cholesterol and desmosterol), plants (dihydrobrassicasterol and 24-methylene cholesterol), algae (fucosterol) and marine invertebrates (clionasterol). In both species, sterols derive from mevalonate through cyclization of squalene to cycloartenol by cycloartenol synthase. The pathway anticipates synthesis of cholesterol by enzymes of the phytosterol route in plants, as recently reported in Solanaceae. Major divergences stem from reduction of Δ24(28) and Δ24(25) double bonds which, in diatoms, are apparently dependent on sterol reductases of fungi, algae and animals. Phylogenetic comparison revealed a good level of similarity between the sterol biosynthetic genes of S. marinoi and C. cryptica with those in the genomes of the other diatoms sequenced so far

Plant-based beverages as good sources of free and glycosidic plant sterols

To address the ever-growing group of health-conscious consumers, more and more nutritional and health claims are being used on food products. Nevertheless, only very few food constituents, including plant sterols, have been appointed an approved health claim (European Commission and Food and Drugs Administration). Plant sterols are part of those limited lists of approved compounds for their cholesterol-lowering properties but have been praised for their anti-inflammatory and anti-carcinogenic properties as well. Despite this indisputable reputation, direct quantitative data is still lacking for naturally present (conjugated) plant sterols in beverages. This study aimed to fill this gap by applying a validated extraction and UPLC-MS/MS detection method to a diverse range of everyday plant-based beverages. B-sitosterol--D-glucoside (BSSG) showed to be by far the most abundant sterol in all beverages studied, with concentrations up to 60–90 mg per 100 mL in plant-based milk alternatives and fresh fruit juices. Ergosterol (provitamin D2) could be found in beers (0.8–6.1 g per 100 mL, from the yeast) and occasionally in juices (17–29 g per 100 mL). Overall, the results demonstrated that the concentrations of water-soluble sterol conjugates have been underestimated significantly and that specific plant-based beverages can be good, low-fat sources of these plant sterols

Localization of sterols and oxysterols in mouse brain reveals distinct spatial cholesterol metabolism

Dysregulated cholesterol metabolism is implicated in a number of neurological disorders. Many sterols, including cholesterol and its precursors and metabolites, are biologically active and important for proper brain function. However, spatial cholesterol metabolism in brain and the resulting sterol distributions are poorly defined. To better understand cholesterol metabolism in situ across the complex functional regions of brain, we have developed on-tissue enzyme-assisted derivatization in combination with microliquid extraction for surface analysis and liquid chromatography-mass spectrometry to locate sterols in tissue slices (10 µm) of mouse brain. The method provides sterolomic analysis at 400-µm spot diameter with a limit of quantification of 0.01 ng/mm2. It overcomes the limitations of previous mass spectrometry imaging techniques in analysis of low-abundance and difficult-to-ionize sterol molecules, allowing isomer differentiation and structure identification. Here we demonstrate the spatial distribution and quantification of multiple sterols involved in cholesterol metabolic pathways in wild-type and cholesterol 24S-hydroxylase knockout mouse brain. The technology described provides a powerful tool for future studies of spatial cholesterol metabolism in healthy and diseased tissues

Effects of genotype and sowing date on phytostanol-phytosterol content and agronomic traits in wheat under organic agriculture

Cereals are an important source of sterols and stanols in the human diet. The present study underlines the effect of genotype and weather conditions in bread wheat, on total sterol and stanol content (TSS), agronomic traits, proteins and ash content under organic conditions. Variations in TSS as well as other characters between two sowing dates were observed. A broad genotypic variability was also reported since extreme genotypes differed by more than 30 mg 100. g-1 DW for TSS, with total stanol content varying twofold. Moreover, two groups of genotypes that differed in agronomic production, ash and protein content were depicted, based on their response to an increase in temperature. This result suggests that the genotypic factor prevails over the sowing date factor for determining sterol and stanol traits in wheat cultivated under organic conditions. Nevertheless, a strong interaction exists between the two factors, which can be used to drive bioaccumulation of these molecules

Olive oil

Analyses of phytosterol classes of olive and hazelnut oils collected from different countries by TLC, GC and GC-MS revealed considerable quantitative differences. The composition of 4-desmethyl- and 4-monomethylsterols was similar in both oils, but 4,4'-dimethylsterols composition differed. Lupeol and an unknown (lupane skeleton) compound were exclusively present in hazelnut oil 4,4´-dimethylsterols and could be used as markers to detect virgin olive oil adulteration with hazelnut oil at levels below 4%. Conventional TLC to separate phytosterol classes has a low recovery rate and is time-consuming. A new SPE method to separate phytosterol classes was developed with stepwise elution by increasing the polarity of the n-hexane:diethyl ether solvent mixture. Comparison of the results obtained for hazelnut and virgin olive oils with those of TLC revealed that the SPE method was faster and gave higher sterol recovery rates. Free and esterified forms of sterols provide detailed information on the identity and quality of vegetable oils, and therefore 4,4´-dimethylsterols were investigated in hazelnut oil and virgin olive oil. A sample of solvent-extracted hazelnut oil was refined to monitor the effects of processing on 4,4´-dimethylsterol levels and on specific marker compounds. Of the refining processes tested, only neutralisation and bleaching considerably reduced 4,4´-dimethylsterols. In fully-refined hazelnut oil, losses of marker compounds in free form were higher than losses in their esterified form. GC-MS analysis showed that adulteration of olive oil with fully-refined hazelnut oil could be detected at levels of 2% by tracing lupeol in total/esterified forms of 4,4´-dimethylsterols. Olive oil has many applications in the food industry, e.g. blended with oils such as palm stearin to produce margarine or shortening by chemical interesterification. Investigation on lipid and minor lipid components of an olive oil-palm stearin blend during chemical interesterification showed that sterols were esterified with fatty acids at a higher level at 120 °C (7%) than at 90 °C (4%). Despite heat treatment and several steps to produce an interesterified product, there were minor losses in phytosterol and tocopherol contents and no significant increases in phytosterol oxidation

Cultivar and Year-to-Year Variation of Phytosterol Content in Rye (Secale cereale L.)

Intake of phytosterols (and -stanols) has been shown to decrease the level of low-density lipoprotein cholesterol and thus protect against development of cardiovascular diseases. Therefore, studies on the cultivar and year-to-year variation in phytosterol content in rye grains have been performed. The phytosterol content and composition of different rye cultivars, grown under identical conditions on the same field in three consecutive years, were analyzed. Both cultivar and year-to-year variation in sterol content were statistically significant (p < 0.0001). The total sterol content varied from 1007 ± 21 mg/kg in the highest yielding cultivar, Tsulpan 3, to 761 ± 10 mg/kg in the lowest yielding cultivar (Amando in the 1999 harvest). Because the meteorological conditions varied substantially between the different years, it was possible to deduce the impact of varying weather conditions on phytosterol content in the different cultivars. The studied cultivars had all the lowest phytosterol contents in the dry and warm harvest season of 1999. Although there were statistically significant cultivar and year-to-year variations in the sterol composition (p < 0.0001), these were only between 2 and 4% of the total sterol content

POTENTIAL HEALTHCARE SAVINGS FROM PLANT STEROL ENRICHED FOODS IN CANADA

Increased consumption of foods containing plant sterols has the potential to reduce the incidence of coronary heart disease (CHD) and thus reduce costs associated with treating that disease in a significant way. This paper reports the results of an investigation of the potential monetary benefits of allowing foods enriched with plant sterols to be marketed in Canada. The objective of this research was to estimate the annual savings that would accrue to Canada’s single-payer publicly funded health care system if plant sterols were approved for use. If foods containing plant sterols are consumed at a sufficient rate, a reduction in CHD should follow. This research employs a variation of traditional cost-of-illness analysis entailing four steps: (i) estimation of a “success rate” (proportion of persons who would consume plant sterols at the necessary rate); (ii) presumption of blood cholesterol reduction due to plant sterol consumption; (iii) assumption of reduction in CHD that follows from blood cholesterol reduction; (iv) calculation of cost savings associated with reduced incidence of CHD. Calculations were carried out for four scenarios: ideal, optimistic, pessimistic, and very pessimistic. It was estimated that between $38 million (very pessimistic scenario) and$2.45 billion (ideal scenario) could be saved annually by Canada’s health care system with plant sterol enriched food products being made available for sale.coronary heart disease, cost of illness analysis, health care costs, success rate, Agricultural and Food Policy, Consumer/Household Economics, Demand and Price Analysis, Food Consumption/Nutrition/Food Safety, Food Security and Poverty, Health Economics and Policy, I18,

Microbial community dynamics during composting of sewage sludge and straw studied through phospholipid and neutral lipid analysis

The composting process involves a succession of different communities of microorganisms that decompose the initial material, transforming it into a stable final product. In thiswork, the levels of phospholipid fatty acid (PLFA), neutral lipid fatty acid (NLFA) and sterolwere monitored in compost versus time, as indicators of the activity of various microorganisms (Gram-positive or Gram-negative bacteria, fungi, etc.). During composting, the PLFA and NLFA from Gram-negative bacteria and eukaryotes (2-OH 10; 3-OH 12; 2-OH 14; 13:0; 16:1; 18:1 trans) aswell as some sterols of plant origin (e.g. monostearin sterols) decreased until the end of composting. In contrast, the branched fatty acids with iso- and anteiso-forms (i-15:0; a-15:0; i-16; i-17) increased mainly in the thermophilic phase, but decreased right after. The PLFA 18:2 (6;9), which is used as an index of the occurrence of some fungi, rose strongly at the beginning of composting, but fell after peak heating. In contrast, the other main sterol indicative of fungi, ergosterol, decreased at the beginning of the thermophilic phase, but increased strongly by the end of composting. Accordingly, cluster and PCA analysis separated the PLFA of Gram-negative bacteria and eukaryotic cells from those of Gram-positive bacteria and long-chain fatty acids. The fungal PLFA considered, 18:2 (9, 12), was clustered more closely to iso- and anteiso-branched PLFAs. Stigmasterol, squalene and cholesterol occurred in the lower right part of the loading plot and were clustered more closely to iso-, anteiso-branched PLFAs and 18:2w6,9 suggesting their relationship to microbial activities. We also observed the tendency of resistance of fatty acid PLFAs and NLFAs of long chain (19:0 (cis-9); 20:0) and some recalcitrant sterols, e.g. sitosterol, at the end of composting. The presence of high levels of the latter in the final stage indicates their contribution to the structural stability of organic matter fractions. These recalcitrant components were more clustered and occurred in the lower right part of the loading plot

Sterol concentration and distribution in sunflower seeds (Helianthus annuus L.) during seed development

Sunflower seeds are currently used for edible oil production. Among oil minor compounds, phytosterols are of special interest due to their cholesterol reducing properties. This paper reports studies on their accumulation and distribution in the embryo and hull, and the effects of temperature on phytosterol contents in sunflower seed produced under both conventional and organic field conditions. An optimized method of sterol determination, adapted to studies on small samples of seed, is presented. Seventy-two % of phytosterols were found in the embryo, 28 % in the hull. The periods of phytosterols concentration varied according to sterol category and seed part. Application of these results to improve production of natural sterols for functional food use is discussed