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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., (© 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

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 (IP(6)). 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 IP(6) 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 IP(6) in 48 h; 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 IP(6) was degraded after 48 h. This corresponds to a remaining level of 0.4 and 0.3μmol IP(6)/g dw. Co-inoculation with L. plantarum did not increase IP(6) degradation. Moreover, fermentation with P. kudriavzevii TY13 yielded a successive increase in inorganic phosphate (P(i)), from 0.7 to 5.4 mM, suggesting a phytase production in TY13 which is fairly insensitive to P(i) 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 © 2011 Elsevier B.V. All rights reserved.)

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.

Using a combination of High-Performance Ion Chromatography analysis and kinetic studies, the pathway of myo-inositol hexakisphosphate dephosphorylation by a phytase from a Malaysian waste-water bacterium was established. The data demonstrate that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-I(1,2,3,4,5)P(5), D-I(2,3,4,5)P(4), D-I(2,3,4)P(3), D-I(2,3)P(2) to finally I(2)P. It was estimated that more than 90% of phytate hydrolysis occurs via D-I(1,2,3,4,5)P(5). Thus, the phytase from the Malaysian waste-water bacterium has to be considered a 6-phytase (E.C. 3.1.3.26). A second pathway of minor importance could be proposed which is in accordance with the results obtained from analysis of the dephosphorylation products formed by the action of the phytase under investigation on myo-inositol hexakisphosphate. It proceeds via D/L-I(1,2,4,5,6)P(5), D/L-I(1,2,4,5)P(4), D/L-I(1,2,4)P(3), D/L-I(2,4)P(2) to finally I(2)P.

Using a combination of high-performance ion chromatography analysis and kinetic studies, the pathway of myo-inositol hexakisphosphate dephosphorylation by the beta-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 beta-propeller phytases. The data demonstrate that all of these beta-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 beta-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).

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.

Using a combination of high-performance ion chromatography analysis and kinetic studies, the pathway of dephosphorylation of myo-inositol hexakisphosphate by the phytases purified from faba bean and lupine seeds, respectively, was established. The data demonstrate that the legume seed phytases under investigation dephosphorylate myo-inositol hexakisphosphate in a stereospecific way. The phytase from faba bean seeds and the phytase LP2 from lupine seeds degrade phytate by sequential removal of phosphate groups via D-Ins(1,2,3,5,6)P(5), D-Ins(1,2,5,6)P(4), D-Ins(1,2,6)P(3), and D-Ins(1,2)P(2) to finally Ins(2)P, whereas the phytases LP11 and LP12 from lupine seeds generate the final degradation product Ins(2)P via D-Ins(1,2,4,5,6)P(5), D-Ins(1,2,5,6)P(4), D-Ins(1,2,6)P(3), and D-Ins(1,2)P(2).

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)P5 to Ins(2,4,5,6)P4 to finally Ins(2,4,6)P3 or D-Ins(2,5,6)P3 and via D-Ins(1,2,4,5,6)P5 to D-Ins(1,2,5,6)P4 to finally D-Ins(1,2,6)P3. The resulting myo-inositol trisphosphate D-Ins(1,2,6)P3 was degraded via D-Ins(2,6)P2 to finally Ins(2)P after prolonged incubation times in combination with increased enzyme concentration.

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]

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]

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]

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]