Ann-Sofie Sandberg

Bioavailability of minerals in legumes

The mineral content of legumes is generally high, but the bioavailability is poor due to the presence of phytate, which is a main inhibitor of Fe and Zn absorption. Some legumes also contain considerable amounts of Fe-binding polyphenols inhibiting Fe absorption. Furthermore, soya protein per se has an inhibiting effect on Fe absorption. Efficient removal of phytate, and probably also polyphenols, can be obtained by enzymatic degradation during food processing, either by increasing the activity of the naturally occurring plant phytases and polyphenol degrading enzymes, or by addition of enzyme preparations. Biological food processing techniques that increase the activity of the native enzymes are soaking, germination, hydrothermal treatment and fermentation. Food processing can be optimized towards highest phytate degradation provided that the optimal conditions for phytase activity in the plant is known. In contrast to cereals, some legumes have highest phytate degradation at neutral or alkaline pH. Addition of microbial enzyme preparations seems to be the most efficient for complete degradation during processing. Fe and Zn absorption have been shown to be low from legume-based diets. It has also been demonstrated that nutritional Fe deficiency reaches its greatest prevalence in populations subsisting on cereal- and legume-based diets. However, in a balanced diet containing animal protein a high intake of legumes is not considered a risk in terms of mineral supply. Furthermore, once phytate, and in certain legumes polyphenols, is degraded, legumes would become good sources of Fe and Zn as the content of these minerals is high.

The usefulness of in vitro models to predict the bioavailability of iron and zinc: A consensus statement from the HarvestPlus expert consultation

A combination of dietary and host-related factors determines iron and zinc absorption, and several in vitro methods have been developed as preliminary screening tools for assessing bioavailability. An expert committee has reviewed evidence for their usefulness and reached a consensus. Dialyzability (with and without simulated digestion) gives some useful information but cannot predict the correct magnitude of response and may sometimes predict the wrong direction of response. Caco-2 cell systems (with and without simulated digestion) have been developed for iron availability, but the magnitude of different effects does not always agree with results obtained in human volunteers, and the data for zinc are too limited to draw conclusions about the validity of the method. Caco-2 methodologies vary significantly between laboratories and require experienced technicians and good quality cell culture facilities to obtain reproducible results. Algorithms can provide semi-quantitative information enabling diets to be classified as high, moderate, or low bioavailability. While in vitro methods can be used to generate ideas and develop hypotheses, they cannot be used alone for important decisions concerning food fortification policy, selection of varieties for plant breeding programs, or for new product development in the food industry. Ultimately human studies are required for such determinations.

Phytate degradation during breadmaking: Effect of phytase addition

Bread was made using whole wheat flour and flour of 60% extraction, and inositol hexaphosphate and its hydrolysis products were measured by high-performance liquid chromatography (HPLC) in both doughs and breads. Addition of a phytase preparation from A. niger to the doughs resulted in an increased degradation of phytate. When milk was included in the dough formulation, phytate degradation was inhibited almost completely, but fermented milk had no effect. Lactic acid, whether in the presence or absence of calcium chloride, inhibited phytate degradation, although when phytase was added phytate hydrolysis was complete.

The effect of wheat bran on the absorption of minerals in the small intestine

1. Studies on mineral absorption were carried out in ileostomy patients using the metabolic balance technique. The effect of wheat bran on the absorption of phosphorus, calcium, magnesium, zinc and iron was studied. The extent of digestion of bran phytate in the stomach and small intestine was also investigated. 2. Eight patients with well established conventional ileostomies were studied during two periods while on a constant low-fibre diet. In the second period, 16 g wheat bran/d (American Association of Cereal Chemists) was added to the diet. The amount of phytate-P, non-phytate-P, Ca, Mg, Zn and Fe was determined in the ileostomy contents and in duplicate portions of the diet. 3. Of the added bran phytate-P 24-61% was recovered in the ileostomy contents. In the bran period a significantly decreased amount of Zn was absorbed, while the apparent absorption of Fe and phytate-P increased and that of non-phytate-P, Ca and Mg remained constant. Due to the mineral content of bran, the relative absorption differed in some respects from the absolute absorption, being decreased for Zn, Mg and phytate-P but unchanged for Ca, Fe and non-phytate-P. 4. It is concluded that phytate is partly digested in the stomach and small intestine or possibly absorbed. Addition of 16 g bran/d to the diet does not seem to impair the mineral absorption from the small intestine except that of Zn.

Improved zinc and iron absorption from breakfast meals containing malted oats with reduced phytate content

The absorption of Zn or Fe from breakfast meals containing oat porridge prepared from malted and soaked oats and a control porridge made from untreated oats was measured in human subjects. The effect on Zn and Fe absorption of reducing the phytate content of oat-porridge meals was examined in each subject by extrinsic labelling of porridge with 65Zn and of bread rolls with 55Fe and 59Fe, and measuring whole-body retention and the erythrocyte uptake of isotopes. Each experiment comprised nine to ten subjects. The absorption of Zn from malted-oat porridge with a phytate (inositol hexaphosphate) content of 107 mumol was 18.3%, and significantly higher (P < 0.05) than from the control porridge containing 432 mumol phytate (11.8%). Fe absorption from the meal containing malted-oat porridge with 107 mumol phytate (Expt 2) was also significantly improved (P < 0.05) compared with that from the meal containing control porridge with 437 mumol phytate. The average increase in Fe absorption was 47%, or from 4.4 to 6.0%. In the breakfast meal containing malted porridge with 198 mumol phytate (Expt 3) the increase in Fe absorption was not significantly improved. Even though the phytate content was reduced to a greater extent in Expt 3 than Expt 2, the average increase in Fe absorption in Expt 3 was only 25% more than that from the meal containing control porridge (with 599 mumol phytate), depending on the higher absolute amount of phytate. In conclusion, an improvement in Zn and Fe absorption from oat products can be achieved by practising malting and soaking in the processing of oats. This may be of importance in the prevention of mineral deficiency in vulnerable groups.

The effect of food processing on phytate hydrolysis and availability of iron and zinc

Phytate is one of the major inhibiting factor for zinc and iron absorption. When phytate is hydrolyzed during the food process the mineral availability is increased. By activation of the endogenous enzyme phytase which is present in plant foods, or addition of phytase, phytate is degraded to various inositolphosphates containing 1-5 phosphate groups per an inositol molecule. The effects of degradation products of phytate on availability of zinc, calcium and iron have to be further investigated. Food processes including soaking, germination and fermentation were under optimal conditions demonstrated to completely reduce the phytate content of cereals and vegetables. The results were related to in vitro measurements of iron availability and human iron and zinc absorption studies.

Phytate reduction in bread containing oat flour, oat bran or rye bran

Phytate reduction in bread baked with oat flour and in bread with rye bran or oat bran addition was examined in scalded bread and bread made with varying amounts of sour dough. A considerable decrease in phytate content was observed in bread containing rye bran, ranging from 66-97 % of the initial phytate content in the raw materials. The most marked phytate reduction, of 96-97 %, occurred in bread made with 10 % sour dough (pH 4·6) or in bread in which the pH had been adjusted in the scalding with lactic acid, resulting in a pH between 4·4 and 5·1 in both dough and bread. In absolute amounts, more phytate was hydrolysed in the scalded breads with pH between 4·4 and 5·4 than in breads made with sour dough. The most effective phytate decomposition in bread with oat flour or oat bran occurred in bread made from scalded oat flour and which had a sour dough content of 20 or 30 %. In these breads, with pH values between 4·3 and 4·6 in dough and breads, the phytate was reduced from the initial content by 96%. The phytate content in unsoured bread with scalded oat bran was reduced at most to 62% of the initial amount.

Phytate degradation by human gut isolated Bifidobacterium pseudocatenulatum ATCC27919 and its probiotic potential

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.

Processing of quinoa (Chenopodium quinoa, Willd): effects on in vitro iron availability and phytate hydrolysis

The effect of different processing techniques was studied on in vitro iron availability and phytate hydrolysis in high and low saponin content quinoa (Chenopodium quinoa, Willd) seeds. Water slurries of ungerminated and germinated quinoa flour were processed by cooking, soaking, and fermentation using Lactobacillus plantarum as starter. Iron solubility under physiological conditions (in vitro) was measured and used as an estimation of iron availability. Phytate (inositol hexaphosphate/IP6) and its degradation products were analysed by an HPLC method. The IP6 + IP5 content was reduced by cooking with 4 to 8%, germination with 35 to 39%, soaking with 61 to 76% and by fermentation with 82 to 98%. The highest reduction, about 98%, was obtained after fermentation of the germinated flour. Cooking had no effect on the amount of soluble iron. Iron solubility increased, however, two to four times after soaking and germination, three to five times after fermentation and five to eight times after fermentation of the germinated flour samples and was highly correlated to the reduction of IP6 + IP5 (P < 0.001). There was no difference between the quinoa varieties with regard to phytate reduction and iron solubility. The pH in fermented samples was reduced from 6.5 to about 3.5, due to lactic acid formation.

Malting of oats in a pilot-plant process. Effects of heat treatment, storage and soaking conditions on phytate reduction

Abstract Phytate reduction in oats during malting in a pilot-plant process and soaking under optimal conditions for oat phytase was studied to investigate whether the malting process developed in the laboratory was also applicable on a large scale. The effects of different drying and storage conditions were also investigated. In malted oats that had been dried at 30°C and stored frozen and entire for 1·5 months, the phytate content was reduced to 0·2 μmol/g (99% reduction) after soaking as a flour for 17 h at 37·8°C. The same soaking, repeated with oats dried at 80°C and stored as whole grains at +4°C for 1·5 months, reduced the phytate content to 0·3 μmol/g (98% reduction). Prolonged storage (12 months) resulted in slower phytate hydrolysis in oats dried at 80°C (92% reduction) during soaking for 17 h at 37·8°C. Pre-soaking of malted oat flour for 16 h at room temperature was used to achieve pH conditions that were closer to the optimum for phytase activity, thus resulting in more efficient phytate reduction. Of the conditions studied in this report, the optimum pH for oat phytase during soaking was found to be between pH 4 and 4·5. In whole malted grains, the phytate content was reduced at most by 79% after soaking as whole kernels.