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Highlights • A new alternative to classic fishmeal production is proposed, using the pH shift method. • The pH shift process is proposed as an efficient method of working with high bone-low meat sources, such as fishery byproducts. • Protein yield is affected by the treatment that is applied during the pH shift process. • Proximate composition of the pH shift produced protein concentrates is also subject to treatment. Abstract There is a strong need in animal production for sustainable protein sources used in feed. The aim of the present study was to evaluate the potential of the pH-shift process as a method to concentrate proteins from complex bone-rich herring (Clupea harengus) by-products. By-product parts (frame, head, guts), protein solubilization pH (pH 2.8 or 11.3) and separation g-force (800 or 8000 g) were varied. The findings revealed that protein recovery yields of up to 60% were obtained and that higher protein yield was obtained from the alkaline than the acid process. By-product part and g-force had no significant impact on protein yield. Protein levels reached in the concentrates were 75–80 % (dw basis). Protein concentrates contained a similar amino acid profile as have been reported for fishmeal, with ∼40% essential amino acids and up to 7.8% and 2.8% of lysine and methionine. Residual levels of LC n-3 PUFA as well as calcium, magnesium, potassium and phosphorous were also found in the produced protein concentrates which could be an added value. Acid produced concentrates were particularly high in calcium potassium and phosphorous. Provided a careful evaluation, the pH-shift process could become a successful alternative to classic fish meal production for increasing the use of fish by-products for production of protein ingredients for feed. [ABSTRACT FROM AUTHOR]

The content of zinc, iron, calcium and phytate in the 16 most consumed foods from 5 villages in a tropical rural area of Bolivia was analyzed. The food items were selected according to a completed food frequency questionnaire. Minerals were analyzed by atomic absorption and phytates by HPIC chromatography. The molar ratios of phytate:mineral are presented as indication of the mineral bioavailability. Within the analyzed food, quinoa is a potential source of minerals: zinc 3.65, iron 5.40 and calcium 176 mg/100 g; however, it also has the highest content of phytate 2060 mg/100 g. Cereals and legumes showed high concentration of phytates (from 142 to 2070 mg/100 g), roots and tubers have lower concentrations (from 77 to 427 mg/100 g). In general, both phytate contents and molar ratios Phy:Zn (phytate:zinc), Phy:Fe (phytate:iron) and Phy:Ca (phytate:calcium) in most of the analyzed foods were at levels likely to inhibit the absorption of these minerals. Significant positive associations ( p < 0.01) were found between the level of phytate and minerals in food, for zinc ( r = 0.714), iron ( r = 0.650) and calcium ( r = 0.415). The results compared to data from USA or from Bolivia showed some discrepancies, confirming the need for more reliable data for dietary evaluations and interventions. [ABSTRACT FROM AUTHOR]

Determination of microalgaes' fatty acid content is often done with chloroform and methanol according to the Bligh and Dyer extraction, though faster methods exist. A number of comparisons between the Bligh and Dyer and faster methods have resulted in contradicting data, possibly due to differences in algae used and the different versions of the Bligh and Dyer method applied. Here, various forms of direct-transesterification (D-TE) and two-step transesterification (2-TE), including three versions developed in our lab, are compared with the original Bligh and Dyer (Can J Biochem Physiol 37: 911-917, ) extraction and two modifications thereof (Lee et al. J AOAC Int 79:487-492, , and our own acidified version) on microalgae with different cell walls: Isochrysis galbana, Nannochloropsis oculata, and Phaeodactylum tricornutum. In total, fatty acid extracts from 11 methods were separated and quantified by gas chromatography with mass spectrometry. Results show that, for N. oculata and P. tricornutum, methods based on chloroform-methanol underestimated the fatty acid content compared with the 2-TE and D-TE methods, which gave similar results. Moreover, D-TE methods are faster than chloroform-methanol methods and use chemicals that are less toxic. Of the D-TE methods, the ones using hydrochloric acid and sulfuric acid recovered the most fatty acids, while boron trifluoride recovered slightly less. The main qualitative difference between the fatty acids recovered was that the chloroform-methanol methods recovered less saturated fatty acids in P. tricornutum. [ABSTRACT FROM AUTHOR]

Celiac disease is caused by an inappropriate immune response to incompletely digested gluten proteins. We investigated whether synthetic peptides with high affinity to wheat gliadin could be selected with a phage display technique and whether complexes between such peptides and gliadin could sustain gastric and pancreatic digestion. Two synthetic peptides, P61 and P64, were selected because of their high affinity to immobilized gliadin. They were allowed to form complexes with gliadin, whereafter the complexes were subjected to in vitro digestion with gastric and pancreatic enzymes. The digestion products were analyzed with Western blot and RP HPLC. The results showed that both peptides formed stable complexes with intact gliadin and that complexes between gliadin and peptide P64 partly resisted gastrointestinal digestion. The two peptides reduced the binding of serum anti-gliadin IgA antibodies by 12%, and 11.5%, respectively, and the binding of anti-gliadin antibodies of the IgG isotype by 13% and 10%. Thus peptides produced by a phage display technique could interact stably with gliadin partly masking epitopes for antibody binding. A combination of peptides of this kind may be used to block gliadin-immune system interactions. [ABSTRACT FROM AUTHOR]

Abstract: The growing awareness of the relationship between diet and health has led to an increasing demand for food products that support health above and beyond providing basic nutrition. Probiotics are live organisms present in foods, which yield health benefits related to their interactions with the gastrointestinal tract. Phytases are a subgroup of phosphatases that catalyse the desphosphorylation of phytate, which reduces its negative impact on mineral bioavailability, and generates lower inositol phosphates. The aims of this investigation were to (i) study the ability of the probiotic candidate Bifidobacterium pseudocatenulatum to degrade phytate in synthetic medium, to (ii) identify the lower inositol phosphates generated, to (iii) study its survival under conditions mimicking gastrointestinal passage and finally to (iv) assess adhesion of the bacteria to Caco-2 cells. The first steps of InsP 6 degradation by B. pseudocatenulatum phytate-degrading enzyme/s were preferentially initiated at the DL-6-position and 5-position of the myo-inositol ring. It suggests that the main InsP 6 degradation pathway by B. pseudocatenulatum by sequential removal of phosphate groups was D/L-Ins(1,2,3,4,5)P 5 or D/L-Ins(1,2,3,4,6)P 5; D/L-Ins(1,2,3,4)P 4; to finally Ins(1,2,3)P 3 and D/L-Ins(1,2,4)P 3/D/L-Ins(1,3,4)P 3. This human strain also showed a notable tolerance to bile as well as a selective adhesion capacity (adhesion to control surfaces was zero), to human intestinal Caco-2 cells comparable to the commercial probiotic B. lactis. The phytate-degrading activity constitutes a novel metabolic trait which could contribute to the improvement of mineral absorption in the intestine as a nutritional probiotic feature with potential trophic effect in human gut. [Copyright &y& Elsevier]

For the first time a dual pathway for dephosphorylation of myo-inositol hexakisphosphate by a histidine acid phytase was established. The phytate-degrading enzyme of Klebsiella terrigena degrades myo-inositol hexakisphosphate by stepwise dephosphorylation, preferably via D-Ins(1,2,4,5,6)P5, D-Ins(1,2,5,6)P4, D-Ins(1,2,6)P3, D-Ins(1,2)P2 and alternatively via D-Ins(1,2,4,5,6)P5, Ins(2,4,5,6)P4, D-Ins(2,4,5)P3, D-Ins(2,4)P2 to finally Ins(2)P. It was estimated that more than 98% of phytate hydrolysis occurs via D-Ins(1,2,4,5,6)P5. Therefore, the phytate-degrading enzyme from K. terrigena has to be considered a 3-phytase (EC 3.1.3.8). A second dual pathway of minor importance could be proposed that is in accordance with the results obtained by analysis of the dephosphorylation products formed by the action of the phytate-degrading enzyme of K. terrigena on myo-inositol hexakisphosphate. It proceeds preferably via D-Ins(1,2,3,5,6)P5, D-Ins(1,2,3,6)P4, Ins(1,2,3)P3, D-Ins(2,3)P2 and alternatively via D-Ins(1,2,3,5,6)P5, D-Ins(2,3,5,6)P4, D-Ins(2,3,5)P3, D-Ins(2,3)P2 to finally Ins(2)P. D-Ins(2,3,5,6)P4, D-Ins(2,3,5)P3, and D-Ins(2,4)P2 were reported for the first time as intermediates of enzymatic phytate dephosphorylation. A role of the phytate-degrading enzyme from K. terrigena in phytate breakdown could not be ruled out. Because of its cytoplasmatic localization and the suggestions for substrate recognition, D-Ins(1,3,4,5,6)P5 might be the natural substrate of this enzyme and, therefore, may play a role in microbial pathogenesis or cellular myo-inositol phosphate metabolism. [ABSTRACT FROM AUTHOR]

Structural, functional and nutritional properties of protein recovered from brown seaweed, S. latissima with alkaline solubilization/isoelectric precipitation as a function of different post-harvest stabilization methods were studied. The latter included freezing at −20 °C/-80 °C, oven-drying, sun-drying, freeze-drying and ensilaging. Also, the efficacy of freeze/thaw-aided precipitation (F/T) in improving protein recovery of the process was evaluated. The freeze-dried, oven-dried, and −20 °C frozen seaweeds resulted in significantly higher protein yield than the −80°C-frozen, sun-dried and ensiled biomasses. F/T increased protein precipitation and doubled total protein yield. Sun-drying and −20°C-freezing caused extensive protein degradation as revealed by SDS-PAGE and HP-SEC, while oven-drying altered the seaweed protein structure with less α-helices. Functional properties of the seaweed proteins were remarkably affected by stabilization condition and F/T, but nutritional value of the proteins was only dependent on stabilization method. Thus, to efficiently recover seaweed proteins, its post-harvest stabilization condition must be carefully chosen based on the final application of the proteins. Image 1005 • Protein yield was strongly governed by post-harvest stabilization method. • Freeze/thawing doubled total protein yield of the process. • Functional properties of proteins were remarkably affected by stabilization method. • Structure and molecular weight of proteins were governed by stabilization method. [ABSTRACT FROM AUTHOR]

Bread is a staple food in many countries and an important source of iron and zinc. The bioavailability of these minerals is generally low because of the content of phytic acid. Traditional Iranian breads were prepared with flours of different extraction rates, Sangak at 93% and Barbari at 82%. Breads were dephytinised by addition of Aspergillus niger phytase during in vitro digestion. The effect upon iron and zinc bioaccessibility in the Caco-2 cell model was investigated. The cellular uptake of iron and zinc was lower from Sangak, compared to Barbari, despite higher mineral content in Sangak. Dephytinisation of both breads increased iron uptake in the Caco-2 cells (0.65 vs. 1.64 in Sangak and 0.77 vs. 1.97 ng mg−1 protein in Barbari). Zinc uptake increased from 0.98 to 2.8 in Sangak and from 1.4 to 2.9 ng mg−1 protein in Barbari. Thus, dephytinisation substantially improves iron and zinc bioaccessibility. [ABSTRACT FROM AUTHOR]

Abstract: The fermented cereal-based gruel togwa is used as weaning food for children in Tanzania. Togwa is rich in minerals but these are often not available for uptake in the human intestine due to natural inhibitors, such as phytate (IP6). The yeasts Pichia kudriavzevii TY13, Hanseniaspora guilliermondii TY14 and TY20, isolated from Tanzanian togwa, and selected for high phytase activity in complex yeast medium YPD, were now studied regarding their ability to degrade IP6 in maize-based model togwa. A modified constitutively high-phytase producing Saccharomyces cerevisiae BY80 and commercial Aspergillus ficuum phytase were included for comparison. In addition, a strain of Lactobacillus plantarum was included in the model-togwa set-up. All yeasts in the study grew and reached final cell density 1.5–2 log units higher than the start value. S. cerevisiae BY80 degraded 85% of the IP6 in 48h; the same degradation level as with A. ficuum phytase (89%). Of the togwa-isolated yeasts, P. kudriavzevii TY13 and H. guilliermondii TY14 showed strong phytate degradation in the model-togwa; 95% or more of the initial IP6 was degraded after 48h. This corresponds to a remaining level of 0.4 and 0.3μmol IP6/g dw. Co-inoculation with L. plantarum did not increase IP6 degradation. Moreover, fermentation with P. kudriavzevii TY13 yielded a successive increase in inorganic phosphate (Pi), from 0.7 to 5.4mM, suggesting a phytase production in TY13 which is fairly insensitive to Pi repression. The study shows that phytate in a model togwa is available for yeast phytase enzymes, and addresses the importance of strain selection for effectively degrading the phytate. Certain yeasts originating from togwa seem to have developed a natural high phytase production, and P. kudriavzevii TY13 and H. guilliermondii TY14 seem particularly well adapted to phytate degradation in togwa, and is our choice for further studies and strain improvement. [Copyright &y& Elsevier]

Using a combination of high-performance ion chromatography analysis and kinetic studies, the pathway of myo-inositol hexakisphosphate dephosphorylation by the β-propeller phytase of Shewanella oneidensis was established, which was then compared with that of Bacillus subtilis 168, Bacillus amyloliquefaciens ATCC 15841, and B. amyloliquefaciens 45 β-propeller phytases. The data demonstrate that all of these β-propeller phytases dephosphorylate myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via d-Ins(1,2,4,5,6)P5, Ins(2,4,5,6)P4 to finally Ins(2,4,6)P3. Thus, the β-propeller phytases prefer the hydrolysis of every second phosphate over that of adjacent ones. This finding does not support previous phytate degradation models proposed by J. Kerovuo, J. Rouvinen, and F. Hatzack (2000. Biochem. J. 352: 623–628) and R. Greiner, A. Farouk, M. Larsson Alminger, and N.G. Carlsson (2002. Can. J. Microbiol. 48: 986–994), but seems to fit with the structural model given by S. Shin, N.C. Ha, B.C. Oh, T.K. Oh, and B.H. Oh (2001. Structure, 9: 851–858). [ABSTRACT FROM AUTHOR]

Background In many developed countries, magnesium and calcium intakes do not reach recommendations for a large part of the population. Mineral water may be a useful alternative source of dietary minerals, especially in groups of people at risk for developing deficiency due to low intakes. Aim To assess if the addition of a natural and mineral-rich water increased small bowel mineral absorption in people with ileostomy. Methods A controlled randomised crossover study with two periods of two days each and a minimum 5 days of washout was performed in six ileostomy subjects. Apparent mineral absorption from 0.5 L of natural mineral water with either a high or a low mineral content consumed in the fasting state was compared. The daily addition of minerals corresponded to 2.3 mmol magnesium, 6.9 mmol calcium and 7.7 mmol sulphate. Ileostomy effluents were sampled and analysed for magnesium, calcium and total sulphate. Results When compared with the control, the median absorbed amount of magnesium increased from 0.8 (0–1.34) mmol/d to 1.2 (0.8–1.9) mmol/d, which corresponded to a 30 % increase (P=0.028). Median amount of calcium absorbed increased from 8.3 (6.7–13.6) mmol/d to 14.8 (8.3–20.4) mmol/d, i. e. a 45% increase (P=0.027). The sulphate absorption increased from 1.9 (1.3–2.2) mmol/d to 5.1 (4.2–6.8) mmol/d, which corresponded to 197% increase (P=0.028). Conclusions The mineral-rich water increased absorption of both magnesium and calcium and can therefore be used as an additional source of minerals. However, consumption with meals may be preferred. [ABSTRACT FROM AUTHOR]

Background: Quinoa is a pseudocereal with relatively high content of proteins and minerals that also contains mineral inhibitors such as phytate. The aim of the present study was to evaluate lactic acid fermentation and dry roasting on the nutritional quality and sensory attributes of quinoa. Various processes were evaluated, and quinoa grains were dry‐roasted, milled, and fermented, either with or without the addition of wheat phytase or activated quinoa phytase (added as back‐slop starter), for 10 hr. In other processes, raw quinoa flour was fermented for 10 hr or 4 hr and dry‐roasted. Hedonic sensory evaluation was then performed to evaluate the acceptability of the fermented flours prepared as porridges. Results: The combined dry roasting and fermentation processes significantly (p <.05) degraded phytate between 30% and 73% from initial content. The most effective process was fermentation of raw quinoa flour followed by dry roasting, which improved the estimated zinc and iron bioavailability. Particularly, estimated zinc bioavailability improved from low (Phy:Zn 25.4, Phy·Zn:Ca 295) to moderate (Phy:Zn 7.14, Phy·Zn:Ca 81.5). Phytate degradation was mainly attributed to the activation of endogenous phytase during fermentation. Dry roasting was effective in improving the sensory attributes of the fermented quinoa flour. Porridge made with raw quinoa flour fermented for 4 hr and dry‐roasted was more favorable to overall acceptability than that which was fermented for 10 hr and dry‐roasted. Conclusion: Fermentation of quinoa flour for 4 hr followed by dry roasting was successful in improving both nutritional and sensory attributes of the final product. [ABSTRACT FROM AUTHOR]

BACKGROUND: Pseudocereals are nutrient‐rich grains with high mineral content but also phytate content. Phytate is a mineral absorption inhibitor. The study's aim was to evaluate phytate degradation during spontaneous fermentation and during Lactobacillus plantarum 299v® fermentation of quinoa, canihua, and amaranth grains and flours. It also aimed to evaluate the accessibility of iron, zinc, and calcium and to estimate their bioavailability before and after the fermentation of flours with starter culture. Lactic acid, pH, phytate, and mineral content were analyzed during fermentation. RESULTS: Higher phytate degradation was found during the fermentation of flours (64–93%) than during that of grains (12–51%). Results suggest that phytate degradation was mainly due to endogenous phytase activity in different pseudocereals rather than the phytase produced by added microorganisms. The addition of Lactobacillus plantarum 299v® resulted in a higher level of lactic acid (76.8–82.4 g kg−1 DM) during fermentation, and a relatively quicker reduction in pH to 4 than in spontaneous fermentation. Mineral accessibility was increased (1.7–4.6‐fold) and phytate : mineral molar ratios were reduced (1.5–4.2‐fold) in agreement with phytate degradation (1.8–4.2‐fold) in fermented flours. The reduced molar ratios were still above the threshold value for the improved estimated mineral bioavailability of mainly iron. CONCLUSION: Fermentation proved to be effective for degrading phytate in pseudocereal flours, but less so in grains. Fermentation with Lactobacillus plantarum 299v® improved mineral accessibility and estimated bioavailability in flours. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The demand for vegetable proteins increases globally and seaweeds are considered novel and promising protein sources. However, the tough polysaccharide-rich cell walls and the abundance of polyphenols reduce the extractability and digestibility of seaweed proteins. Therefore, food grade, scalable, and environmentally friendly protein extraction techniques are required. To date, little work has been carried out on developing such methods taking into consideration the structural differences between seaweed species. In this work, three different protein extraction methods were applied to three Swedish seaweeds (Porphyra umbilicalis, Ulva lactuca, and Saccharina latissima). These methods included (I) a traditional method using sonication in water and subsequent ammonium sulfate-induced protein precipitation, (II) the pH-shift protein extraction method using alkaline protein solubilization followed by isoelectric precipitation, and (III) the accelerated solvent extraction (ASE®) method where proteins are extracted after pre-removal of lipids and phlorotannins. The highest protein yields were achieved using the pH-shift method applied to P. umbilicalis (22.6 ± 7.3%) and S. latissima (25.1 ± 0.9%). The traditional method resulted in the greatest protein yield when applied to U. lactuca (19.6 ± 0.8%). However, the protein concentration in the produced extracts was highest for all three species using the pH-shift method (71.0 ± 3.7%, 51.2 ± 2.1%, and 40.7 ± 0.5% for P. umbilicalis, U. lactuca, and S. latissima, respectively). In addition, the pH-shift method was found to concentrate the fatty acids in U. lactuca and S. latissima by 2.2 and 1.6 times, respectively. The pH-shift method can therefore be considered a promising strategy for producing seaweed protein ingredients for use in food and feed. [ABSTRACT FROM AUTHOR]

Quality changes of vacuum-packed Atlantic mackerel (Scomber scombrus) fillets during 12 months’ frozen storage at −27°C and 9 days’ chilled storage at +4°C were evaluated. Freezing at −27°C preserved the long chain n-3 polyunsaturated fatty acids (LC n-3 PUFAs), both in light and dark muscle, vitamin D, and the low molecular weight metabolites (LMW) (studied by high resolution nuclear magnetic resonance spectroscopy, HR NMR). Protein oxidation took place, especially between 1 and 7 months, decreasing water holding capacity and protein extractability. During chilled storage, no lipid or protein oxidation was observed, but lipolysis increased, and several LMW metabolites relevant for sensory and nutritional quality degraded into non-favorable compounds. The content of biogenic amines was high at day 9 (e.g., 18 mg histamine/100 g), jeopardizing safety. Preservation of mackerel fillets by freezing at −27°C is thus a better option compared to prolonged chilled storage at +4°C; the quality was well preserved for 12 months’ frozen storage. [ABSTRACT FROM AUTHOR]

The study was conducted to examine if crude enzymes (E) produced from Aspergillus niger by solid state fermentation could bio-transform carbohydrates, proteins, inositol phosphate (InsP) and saponins in soybean meal (SBM) under low moisture conditions. Two experiments were performed to determine the effect of mixing, moisture content, treatment time and E levels during enzyme hydrolysis. 1) A single replicate 2 3 full factorial screening design was used to determine the effect of the independent continuous variables time (30 and 70 min), moisture (350 and 450 g kg −1 DM), and the category variable mixing (i.e. effect of static conditions vs. active mixing (X3, S/M)) during the enzyme hydrolysis. 2) A three factorial central composite design (CCD) was used to study the effects of moisture content (316–484 g kg −1 DM), time of hydrolysis (16.4–83.6 min), and E content (0.32-3.68 g kg −1 SBM dry matter (DM)). The first experiment demonstrated that hydrolysis of InsP 6 was significantly affected by time (P < 0.01), moisture content (P < 0.0001) and static vs. mixing (P < 0.01). The hydrolysis of Bb-DDMP was also significantly affected by static vs. mixing (P < 0.05). The second experiment demonstrated that the hydrolysis of InsP 6 was significantly affected by time (P < 0.001), moisture (P ˂ 0.001) and enzyme content (P ˂ 0.01). The InsP 6 content was reduced 84% after 70 min, at a moisture content of 450 g kg −1 DM with the addition of 3 g enzyme kg −1 SBM DM, the InsP 5 -InsP 3 were not accumulated at these parameter settings. The hydrolysis of the saponin group Bb was explained by the enzyme square regressor (P < 0.001) and the interaction between moisture and enzyme (P < 0.001) reaching a maximum by the addition of 2.5 g enzymes kg −1 SBM DM at 484 g kg −1 moisture content. No significant effects on soluble protein and dietary fiber were observed. Mink were fed three diets containing 50% of the protein from SBM, SBM treated without enzymes or treated with enzymes. There was a trend of improved ash apparent digestibility in mink by the enzyme treatment (P = 0.07), but not for phosphorous. The apparent digestibility of protein and amino acids (AA) in mink was improved by the heat treatment used to inactivate the enzymes (P < 0.001), however, not by the enzyme treatment itself. [ABSTRACT FROM AUTHOR]

A bottleneck of crustacean larval culture concerns nutrition and associated cannibalism in communal rearing systems, which impact on larval survival, development and growth. For early-stage European lobster, Homarus gammarus larvae, feeding ecology and body composition are largely unknown. We initiated four progressive feeding experiments (novel feed types, feeding regime and feed size and cannibalism effects) on growth and survival, to inform and update husbandry protocols. Performance of larvae offered a dry commercial feed was not significantly different compared with a conventional wet plankton feed of the same ration and size grade (both within 600-1,000 μm). Further experiments found that the same ration of dry feed offered six times daily improved development and growth, over the conventional regime of three times daily. Small-grade dry feed (particles: 250-360 μm) improved larval performance compared with a larger feed (360-650 μm). Larvae were also fed different proportions of dry feed and/or conspecifics in both communal and individual rearing systems (the latter preventing cannibalism via segregation). Individually reared larvae, fed only dead conspecifics, displayed the greatest survival (80%) to postlarvae. This underlines the impact of cannibalism on survival and nutrition in H. gammarus larviculture. A final experiment analysed H. gammarus zoea 1 composition, identifying deficiencies in ash and carbohydrate in lobster feeds. This suggests a need for a species-specific, formulated dry feed for H. gammarus larviculture. Our research represents the first investigation of H. gammarus larval composition and dietary requirements and highlights decreased growth potential associated with providing nutrition solely from generic commercial feed. [ABSTRACT FROM AUTHOR]

Circulating concentrations of vitamin D, 25(OH)D, and 1,25(OH)2D are lower in obese than lean individuals, but little is known about the adipose tissue content of these molecules. The aim of this study was to explore the possibility to use time-of-flight secondary ion mass spectrometry (TOF-SIMS) to measure vitamin D and its metabolites in fat tissue in obese and lean subjects. Abdominal subcutaneous adipose tissue (SAT) biopsies were obtained from three lean and three obese women, and paired biopsies SAT and visceral adipose tissue (VAT) were obtained from three obese subjects during gastric bypass surgery. TOF-SIMS was used to measure vitamin D3, 25(OH)D3, and 1,25(OH)2D3 in adipose tissue. We found that vitamin D3, 25(OH)D3, and 1,25(OH)2D3 in adipose tissue can be measured with TOF-SIMS. In adipose tissue, vitamin D3 and its metabolites were located in adipocyte lipid droplets. The content of vitamin D3 (P = 0.006) and 25(OH)D3 (P = 0.018) were lower in SAT in obese compared with lean women. TOF-SIMS has the potential to semi-quantitatively measure vitamin D metabolites in adipose tissue, and offers a possibility to compare vitamin D levels in different depots and groups of individuals. It also gives the opportunity to explore the localization of vitamin D metabolites at a cellular level. [ABSTRACT FROM AUTHOR]

The pathway of dephosphorylation of myo-inositol hexakisphosphate by the phytate-degrading enzymes of Bacillus subtilis 168, Bacillus amyloliquefaciens ATCC 15841, and Bacillus amyloliquefaciens 45 was established using a combination of high-performance ion chromatography analysis and kinetic studies. The data demonstrate that all the Bacillus phytate-degrading enzymes under investigation dephosphorylate myo-inositol hexakisphosphate by sequential removal of phosphate groups via two independent routes; the routes proceed via D-Ins(1,2,4,5,6)P[sub 5] to Ins(2,4,5,6)P[sub 4] to finally Ins(2,4,6)P[sub 3] or D-Ins(2,5,6)P[sub 3] and via D-Ins(1,2,4,5,6)P[sub 5] to D-Ins(1,2,5,6)P[sub 4] to finally D-Ins(1,2,6)P[sub 3] . The resulting myo-inositol trisphosphate D-Ins(1,2,6)P[sub 3] was degraded via D-Ins(2,6)P[sub 2] to finally Ins(2)P after prolonged incubation times in combination with increased enzyme concentration. Key words: Bacillus spp., myo-inositol phosphate isomers, phytase, phytate degradation.Nous avons reconstitué la voie de déphosphorylation du myo-inositol hexakisphosphate par les enzymes dégradant le phytate qu'on retrouve chez Bacillus subtilis 168, Bacillus amyloliquefaciens ATCC 15841, et Bacillus amyloliquefaciens 45 à l'aide d'analyses de chromatographie ionique à haute performance et d'études de cinétique. Les données démontrent que toutes les enzymes de Bacillus étudiées dégradant le phytate pouvaient déphosphoryler le myo-inositol hexakisphosphate par l'enlèvement séquentiel de groupes phosphate et ce, par le biais de deux voies indépendantes: les voie passent par le D-Ins(1,2,4,5,6)P[sub 5] , le Ins(2,4,5,6)P[sub 4] pour aboutir au Ins(2,4,6)P[sub 3] ou au D-Ins(2,5,6)P[sub 3] , et par le D-Ins(1,2,4,5,6)P[sub 5] , le D-Ins(1,2,5,6)P[sub 4] pour aboutir au D-Ins(1,2,6)P[sub 3] . Le myo-inositol trisphosphate D-Ins(1,2,6)P[sub 3] résultant a été dégradé en du Ins(2)P en passant par le D-Ins(2,6)P[sub 2] , à la suite de temps d'incubation prolongés combinés à une concentration plus élevée d'enzyme.Mots clés : Bacillus spp., isomères de myo-inositol phosphate, phytase, dégradation du phytate.[Traduit par la Rédaction] [ABSTRACT FROM AUTHOR]