Differences in phytase activity and phytic acid content between cultivated and Tibetan annual wild barleys

The Qinghai-Tibetan Plateau in China is considered to be one of the original centers of cultivated barley. At present, little is known about the phytase activity (Phy) or phytic acid content (PA) in grains of Tibetan annual wild barley. Phy and PA were determined in grains of 135 wild and 72 cultivated barleys. Phy ranged from 171.3 to 1299.2 U kg(-1) and from 219.9 to 998.2 U kg(-1) for wild and cultivated barleys, respectively. PA and protein contents were much higher in wild barley than in cultivated barley. Tibetan annual wild barley showed a larger genetic diversity in phytase activity and phytic acid and protein contents and is of value for barley breeding. There is no significant correlation between phytase activity and phytic acid or protein content in barley grains, indicating that endogenous phytase activity had little effect on the accumulation of phytic acid.     

Zinc absorption from low phytic acid genotypes of maize (Zea mays L.), Barley (Hordeum vulgare L.), and Rice (Oryza sativa L.) assessed in a suckling rat pup model

Dietary phytic acid is a major causative factor for low Zn bioavailability in many cereal- and legume-based diets. The bioavailability of Zn in seed of low phytic acid (lpa) variants of maize ( Zea mays L.), rice ( Oryza sativa L.), and barley ( Hordeum vulgare L.) was evaluated using a suckling rat pup model. Suckling rat pups (14 days old, n = 6-8/treatment) were fasted for 6 h and intubated with (65)Zn-radiolabeled suspensions prepared using seed produced by either wild-type (normal phytic acid) or lpa genotypes of each cereal. Test solutions were radiolabeled overnight (all genotypes) or immediately prior to intubation (barley genotypes). Pups were killed 6 h postintubation and tissues removed and counted in a gamma counter. Zn absorption was low from wild-type genotypes of maize (21, 33%) and rice (26%), and phytic acid reduction resulted in significantly higher Zn absorption, 47-52 and 35-52%, respectively. Zn absorption from wild-type barley incubated overnight was high (86-91%), and phytate reduction did not improve Zn absorption (84-90%), which is likely due to endogenous phytase activity. When the wild-type barley solutions were prepared immediately before intubation, Zn absorption was significantly lower (63, 78%) than from the lpa cultivars (92, 96%). Variation in seed or flour phenolic acid levels did not affect Zn absorption. Differences in seed Zn levels did not substantially affect Zn absorption. Thus, when phytic acid is abundant in a diet, it has a larger effect on Zn absorption than the level of Zn. Therefore, reducing the phytic acid content of staple cereal grains may contribute to enhancing Zn nutrition of populations consuming these staple foods.     

Moderate decrease of pH by sourdough fermentation is sufficient to reduce phytate content of whole wheat flour through endogenous phytase activity

Whole wheat bread is an important source of minerals but also contains considerable amounts of phytic acid, which is known to impair their absorption. An in vitro trial was performed to assess the effect of a moderate drop of the dough pH (around 5.5) by way of sourdough fermentation or by exogenous organic acid addition on phytate hydrolysis. It was shown that a slight acidification of the dough (pH 5.5) with either sourdough or lactic acid addition allowed a significant phytate breakdown (70% of the initial flour content compared to 40% without any leavening agent or acidification). This result highlights the predominance of wheat phytase activity over sourdough microflora phytase activity during moderate sourdough fermentation and shows that a slight drop of the pH (pH value around 5.5) is sufficient to reduce significantly the phytate content of a wholemeal flour. Mg "bioaccessibility"of whole wheat dough was improved by direct solubilization of the cation and by phytate hydrolysis.     

Spelt (Triticum aestivum ssp. spelta) as a source of breadmaking flours and bran naturally enriched in oleic acid and minerals but not phytic acid

The nutritional value of breadmaking cereal spelt (Triticum aestivum ssp. spelta) is said to be higher than that of common wheat (Triticum aestivum ssp. vulgare), but this traditional view is not substantiated by scientific evidence. In an attempt to clarify this issue, wholemeal and milling fractions (sieved flour, fine bran, and coarse bran) from nine dehulled spelt and five soft winter wheat samples were compared with regard to their lipid, fatty acid, and mineral contents. In addition, tocopherol (a biochemical marker of germ) was measured in all wholemeals, whereas phytic acid and phosphorus levels were determined in fine bran and coarse bran samples after 1 month of storage. Results showed that, on average, spelt wholemeals and milling fractions were higher in lipids and unsaturated fatty acids as compared to wheat, whereas tocopherol content was lower in spelt, suggesting that the higher lipid content of spelt may not be related to a higher germ proportion. Although milling fractionation produced similar proportions of flour and brans in spelt and wheat, it was found that ash, copper, iron, zinc, magnesium, and phosphorus contents were higher in spelt samples, especially in aleurone-rich fine bran and in coarse bran. Even though phosphorus content was higher in spelt than in wheat brans, phytic acid content showed the opposite trend and was 40% lower in spelt versus wheat fine bran, which may suggest that spelt has either a higher endogenous phytase activity or a lower phytic acid content than wheat. The results of this study give important indications on the real nutritional value of spelt compared to wheat. Moreover, they show that the Ca/Fe ratio, combined with that of oleate/palmitate, provides a highly discriminating tool to authenticate spelt from wheat flours and to face the growing issue of spelt flour adulteration. Finally, they suggest that aleurone differences, the nature of which still needs to be investigated, may account for the differential nutrient composition of spelt and wheat.     

Fermented pigeon pea (Cajanus cajan) ingredients in pasta products

Pigeon pea (Cajanus cajan var. aroíto) seeds were fermented in order to remove antinutritional factors and to obtain functional legume flour to be used as pasta ingredients. Fermentation brought about a drastic reduction of alpha-galactosides (82%), phytic acid (48%), and trypsin inhibitor activity (39%). Fermented legume flours presented a notable increase of fat and total soluble available carbohydrates, a slight decrease of protein, dietary fiber, calcium, vitamin B2, vitamin E, and total antioxidant capacity, and a decrease of soluble dietary fiber, Na, K, Mg, and Zn contents. No changes were observed in the level of starch and tannins as a consequence of fermentation. The fermented flour was used as an ingredient to make pasta products in a proportion of 5, 10, and 12%. The supplemented pasta products obtained had longer cooking times, higher cooking water absorptions, higher cooking loss, and higher protein loss in water than control pasta (100% semolina). From sensory evaluations, fortified pasta with 5 and 10% fermented pigeon pea flour had an acceptability score similar to control pasta. Pasta supplemented with 10% fermented pigeon pea flour presented higher levels of protein, fat, dietary fiber, mineral, vitamin E, and Trolox equivalent antioxidant capacity than 100% semolina pasta and similar vitamins B1 and B2 contents. Protein efficiency ratios and true protein digestibility improved (73 and 6%, respectively) after supplementation with 10% fermented pigeon pea flour; therefore, the nutritional value was enhanced.     

Nutritional potential of raw and free alpha-galactosides lupin (Lupinus albus Var. multolupa) seed flours. Effect of phytase treatment on nitrogen and mineral dialyzability

The effect of the removal of alpha-galactosides from Lupinus albus L. var. multolupa on the chemical composition of the prepared flour and the dialyzability of N, total P, Ca, Mg, Fe, Zn, and Mn was studied. The extraction process caused a significant increase in total and insoluble nitrogen contents and decreased the amount of soluble protein nitrogen. However, neither these changes nor treatment with phytase seemed to considerably affect in vitro protein digestibility. Except for Ca and Cu, total mineral contents were significantly reduced by the extraction process. The process also caused a significant reduction in the dialyzability of all the minerals studied except P. The decrease in mineral dialyzability was partially counteracted by phytase treatment in amounts of 250-500 phytase units/kg of lupin flour. In the case of Fe, mineral dialyzability did not differ significantly between the two lupin flours studied with treatment with 500 phytase units/kg. Zinc dialyzability was the most efficiently improved by phytase treatment (P < 0.0001), followed by P, Fe, and Mn, and finally by Ca and Mg (P < 0.05).     

Effect of processing methods on the calcium, phosphorus, and phytic acid contents and nutritive utilization of chickpea (Cicer arietinum L.).

The effect of chickpea (Cicer arietinum L.) processing methods on the nutritive utilization of calcium and phosphorus and on phytic acid, a seed component that affects mineral utilization, was studied. Chemical and biological methods were used for nutritional determinations in growing rats. The digestive utilization of calcium from raw chickpea was adequate for growing rats; however, processing resulted in a slight decrease. The metabolic utilization of chickpea calcium was low because of the low rates of net absorption. This was reflected in the decreased calcium levels in longissimus dorsi muscle in the absence of mobilization of calcium from the femur. Soaking in acid solution followed by cooking decreased phytic acid content, suggesting that processing made part of the phytic acid phosphorus available. The absorbed phosphorus was greater than the nonphytic phosphorus supplied by the diet. The digestive utilization of phosphorus was similar in processed and raw chickpeas, despite the loss of soluble anion as a result of processing. These results may indicate the contribution of phosphorus in the form of inositol hexaphosphate-phosphorus.     

豌豆粉添加对米粉品质特性的影响

为了提高米粉的营养价值,本研究将不同粒径的豌豆粉添加到米粉中,分析不同粒径及添加量(0%、7.5%、15%、30%)对大米粉粉质特性及米粉蒸煮、质构和感官特性的影响。结果表明,添加豌豆粉可以增加米粉中蛋白质的含量,添加30%豌豆粉后米粉的蛋白质含量为原米粉的1.73倍。豌豆粉的添加降低了米粉的峰值黏度、最终黏度和回生值,提高了米粉冷糊稳定性,使米粉不易老化。随着豌豆粉添加量的增加,米粉的硬度与蒸煮损失逐渐增大,感官品质降低。豌豆粉的粒度对米粉品质影响较大,当豌豆粉添加量相同时,添加200目豌豆粉的米粉蒸煮损失比添加80目豌豆粉降低9.08%~20.73%;感官评价总分提高1.35%~10.43%。综上,通过降低豌豆粉的粒度可以制备出豌豆粉添加量为30%的品质较好的米粉。本研究结果为营养健康型米粉的开发提供了一定的理论基础。     

Quantitative analysis of phytate globoids isolated from wheat bran and characterization of their sequential dephosphorylation by wheat phytase

Wheat phytase was purified to investigate the action of the enzyme toward its pure substrate (phytic acid - myo-inositol hexakisphosphate) and its naturally occurring substrate (phytate globoids). Phytate globoids were purified to homogeneity from wheat bran, and their nutritionally relevant parameters were quantified by ICP-MS. The main components of the globoids were phytic acid (40% w/w), protein (46% w/w), and several minerals, in particular, K > Mg > Ca > Fe (in concentration order). Investigation of enzyme kinetics revealed that K(m) and V(max) decreased by 29 and 37%, respectively, when pure phytic acid was replaced with phytate globoids as substrate. Time course degradation of phytic acid or phytate globoids using purified wheat phytase was followed by HPIC identification of inositol phosphates appearing and disappearing as products. In both cases, enzymatic degradation initiated at both the 3- and 6-positions of phytic acid and end products were inositol and phosphate.     

Differences in the Aleurone Layer Fate Between Hard and Soft Common Wheats at Grain Milling

In the milling process, efficient separation between the starchy endosperm and the other grain tissues is a key parameter estimated by ash measurement. Because this separation occurs near the aleurone layer interface, better understanding of this tissue fractionation is critical for a better analysis of the wheat milling behavior. Samples from hard and soft common wheat cultivars that had the same protein content were processed on a pilot mill, and whole grain meals or flour streams were analyzed for ash content. The para‐coumaric acid (p‐CA) and phytic acid flour contents were compared with ash measurement and used as markers of the aleurone cell walls or aleurone cell content, respectively. A greater amount of phytic acid in hard wheat flour compared with soft wheat flour was found and reveals a distinct milling behavior between those wheat classes, mainly at the breaking step. Therefore simple ash content measurement is not sufficient to analyze flour purity. At the reduction stage, quantity of phytic acid increases with the other markers and may result from the overall mechanical resistance of the aleurone tissue. As a consequence, wheat hardness not only determines grain milling behavior but also affects flour composition.     

Characterization of phytase activity from cultivated edible mushrooms and their production substrates

Phytase is used commercially to maximize phytic acid degradation and to decrease phosphorus levels in poultry and swine manure. To determine phytase content in edible mushrooms, basidiomata of Agaricus bisporus and three specialty mushrooms (Grifola frondosa, Lentinula edodes, and Pleurotus cornucopiae) and spent mushroom substrate (SMS) were surveyed. Enzyme activity ranged from 0.046 to 0.074 unit/g of tissue for four A. bisporus types (closed and open whites and closed and open browns) grown at The Pennsylvania State University's Mushroom Test Demonstration Facility (MTDF). The addition of various nutrient supplements to phase II mushroom production substrate did not alter phytase activity in A. bisporus. Portabella mushrooms (open brown) obtained from a commercial farm had significantly higher levels of phytase activity (0.211 unit/g of tissue) compared to A. bisporus grown at the MTDF. Of the specialty mushrooms surveyed, maitake (G. frondosa) had 20% higher phytase activity (0.287 unit/g of tissue) than commercial portabella mushrooms. The yellow oyster mushroom (P. cornucopiae) ranked second in level of phytase activity (0.213 unit/g of tissue). Shiitake (L. edodes) contained the least amount of phytase in basidiomata (0.107 unit/g of tissue). Post-crop steam treatment (60 degrees C, 24 h) of SMS reduced phytase activity from 0.074 to 0.018 unit/g. Phytase was partially purified from commercially grown portabella basidiomata 314-fold with an estimated molecular mass of 531 kDa by gel filtration chromatography. The optimum pH for activity was 5.5, but appreciable phytase activity was observed over the range of pH 5.0-8.0. Partially purified A. bisporus phytase was inactivated following a 10-min incubation at > or =60 degrees C.     

Strains of lactic acid bacteria isolated from sour doughs degrade phytic acid and improve calcium and magnesium solubility from whole wheat flour

Five strains of lactic bacteria have been isolated from sour doughs and examined for their ability to degrade phytic acid. In white flour medium in which phytic acid was the only source of phosphorus, the disappearance of phytate and an elevation of inorganic phosphate were observed after only 2 h of incubation in all strains tested (-30 and +60%, respectively). Both phenomena correspond to phytate breakdown. No difference was observed in the levels of phytic acid hydrolysis among strains, suggesting that phytase enzymes are similar among these bacteria. Using whole wheat flour medium naturally rich in phytic acid in the presence of Leuconostoc mesenteroides strain 38, a 9 h fermentation established that the degradation of PA and the production of lactic acid lead to greater Ca and Mg solubility than in control medium.     

菜籽粕植酸提取和分离蛋白的制备

植酸和蛋白是菜籽粕中2种极具经济价值的成份。为提高菜籽粕的综合利用效果,该文以双低冷榨菜籽粕为原料,采用醋酸溶液提取植酸,在膜分离精制植酸粗提液过程中同时回收蛋白;再对植酸提取后的残余物进行蛋白分离,超滤纯化后获得高纯度的蛋白成品。响应面优化的植酸最适提取条件为：醋酸质量分数0.7%,提取温度48℃,液料比10 mL/g,提取时间1.6 h,该条件下植酸得率为1.865%。植酸粗提液中回收出的蛋白和损失植酸分别占菜籽粕的3.63%和0.395%。超滤精制的分离蛋白可达到70%～90%不同纯度的要求,蛋白中多酚含量显著减少,且植酸与硫苷未检出。     

Phenolics, phytic acid, and phytase in Canadian-grown low-tannin faba bean (Vicia faba L.) genotypes

Thirteen low-tannin faba bean genotypes grown at two locations in north central Alberta in 2009 were evaluated to investigate the variation in seed characteristics, phenolic and phytate contents, and phytase and antioxidant activities and to elucidate the relationship of these components as a breeding strategy. Seed characteristics including color were predominantly genotype dependent. The faba bean genotypes with total phenolic content ranging from 5.5 to 41.8 mg of catechin equiv/g of sample was linearly related to tannin content and the best predictor of antioxidant activity. Phytic acid content and phytase activity varied significantly among genotypes and between locations, ranging from 5.9 to 15.1 g/kg and from 1606 to 2154 FTU/kg sample, respectively. Multivariate data analysis performed on 19 components analyzed in this study using principal component analysis (PCA) and cluster analysis demonstrate that differences in seed characteristics, phenolic components, phytic acid, and phytase are major factors in segregating faba bean genotypes. The relatively low phytic acid content and high phytase activity of these low-tannin faba bean genotypes are beneficial/essential traits for their use in human and animal nutrition.     

氮素和水分处理对稻米植酸含量和蛋白组分的影响

以直立密穗型粳稻品种秀水110和弯曲散穗型粳稻品种春江15为材料,对不同氮素水平处理和水分管理方式下稻米植酸含量和蛋白组分的影响及其互作效应进行了研究。结果表明,旱作栽培处理会导致水稻籽粒中植酸含量上升,而施氮处理对籽粒植酸含量的影响效应与水稻的水分管理方式有关,在常规水作条件下,高氮处理（N3）的籽粒植酸含量有所提高,但在旱作栽培方式下,中氮（N2）和低氮处理（N1,不施N）的籽粒植酸含量却略高于高氮处理（N3）,氮素水平与水分管理方式间的互作效应明显; 水稻籽粒植酸含量与粗蛋白总量、4种蛋白组分（清蛋白、球蛋白、醇溶蛋白和谷蛋白）及有关产量性状指标（有效穗数、每穗粒数、千粒重和结实率）在不同水肥处理间的相关性不显著,但氮肥用量过高不仅会导致水稻的产量水平下降,而且也不利于稻米营养品质的改良; 水肥处理对水稻籽粒植酸含量、蛋白总量和4种蛋白组分穗内粒位分布也存在一定影响,着生在稻穗下部的弱势粒,其稻米植酸含量高于稻穗上部或中部的强势粒,因此改善弱势粒灌浆的水肥管理措施将有利于稻米植酸含量的降低。     

Effect of heat treatment on nutritional quality of germinated legume seeds

The effect of heat treatment of germinated soybean, lupin, and black bean on chemical composition and protein utilization in rats was evaluated. Heat treatment caused complete inactivation of trypsin inhibitors whereas it did not affect phytic acid levels. Proximate components, minerals, and amino acids did not change, but low molecular weight sugars were affected by heat treatment differently for each germinated legume. The sugar digestibility ratio (total digestible sugars/total nondigestible sugars) in germinated black beans doubled after heat treatment. True protein digestibility (TD) increased with heat treatment only in germinated soybean. Net protein utilization was markedly improved (20%) with heat treatment in germinated soybean and lupin. Utilizable protein of heat-treated germinated legumes was 289, 236, and 132 g/kg of legume dry weight for soybean, lupin, and black bean, respectively. Supplementation with methionine did not alter TD but improved all other indices of protein utilization in the germinated legumes, particularly in black bean. All three germinated legumes become equivalent in protein quality when heating and supplementation with methionine are combined with germination. The use of germinated heat-treated soybean, lupin, and black bean on their own and/or as food ingredients is nutritionally advantageous due to the low content of nondigestible oligosaccharides and the high protein utilization.     

Optimal conditions for phytate degradation, estimation of phytase activity, and localization of phytate in barley (cv. Blenheim)

Using a multivariate experimental design, optimal conditions for phytate degradation were found to be pH 4.8 and 57 degrees C in barley flour (cv. Blenheim) and pH 5.2 and 47 degrees C in a crude extracted phytase from barley. Three methods for measuring phytase activity in raw and hydrothermally processed barley were compared. Incubation at pH 5 and 55 degrees C for 60 min did not give significantly different results (p > 0.05), whereas incubation at pH 5 and 50 degrees C for 10, 20, 30, and 60 min gave significantly different results (p < 0.001) between methods. The change in microstructure of phytate globoids during hydrothermal processing showed that the degradation was highest in the scutellum cells and less in the aleurone layer.     

植酸对霍山石斛类原球茎悬浮培养细胞生长和多糖合成的影响

研究了植酸对霍山石斛类原球茎悬浮培养细胞生长、多糖合成和培养基中主要营养成分消耗的影响,并分析了细胞中可溶性还原糖、可溶性蛋白质、丙二醛含量以及过氧化物酶和多酚氧化酶的活性。结果表明,植酸能抑制过氧化物酶和多酚氧化酶的活性,提高细胞活力,从而促进细胞生长和多糖合成,以2.5g/L浓度的植酸效果最好,培养36d时,类原球茎干重为29.4g/L,多糖产量为2.06g/L。动力学分析表明,添加2.5g/L的植酸还有利于碳、氮、磷等营养物质的吸收。     

Purification and characterization of phytase induced in tomato roots under phosphorus-deficient conditions

Phytase (myo-inositol hexakisphosphate phosphohydrolase; EC 3.1.3.8) was purified from roots of tomato plants grown under phosphorus-deficient conditions using five purification schemes. The phytase was successfully separated from the major acid phosphatase to an electrophoretic homogeneity. The native molecular weight of this enzyme was estimated to be about 164 kD by Bio-Gel P-200 gel filtration. The molecular weight of the subunit on SDS-PAGE was approximately 82 kD, indicating that the native form of the enzyme was a homodimer. The isoelectric point of tomato phytase was about 5.5. The enzyme exhibited a high affinity for phytic acid (K _m = 38 μM), and was strongly inhibited by phosphate, molybdate and fluoride. Among other characteristics of tomato phytase, the pH and temperature optima were 4.3 and 45°C, respectively. Tomato phytase contained a fairly high concentration of aspartic, glutamic acid and glycine residues.     

氮肥运筹对寒地水稻籽粒植酸、蛋白质和矿质元素的影响

为探究寒地水稻优质栽培的氮肥施用方式,以粳稻品种垦粳7号和垦粳8号为试验材料,通过2年(2017-2018年)田间试验,设置4种氮肥运筹方式:习惯施肥(T1,施氮量150 kg·hm^-2,基肥:蘖肥:穗肥=6:3:1)、氮肥后移1(T2,施氮量150 kg·hm^-2,基肥:蘖肥:穗肥=4:3:3)、基于T2的减氮施肥(T3,施氮量135 kg·hm^-2,基肥:蘖肥:穗肥=4:3:3)和氮肥后移2(T4,施氮量150 kg·hm^-2,基肥:蘖肥:穗肥=5:3:2),其中以T1为对照,研究氮肥运筹对寒地水稻籽粒植酸、总蛋白质及组分、矿质元素含量的影响及其互作效应。结果表明,氮肥、品种对水稻籽粒中植酸含量的影响均达显著或极显著水平。与T1相比,T2、T3和T4水稻籽粒中植酸含量平均分别显著降低了31.92%、48.94%和21.28%;T2、T3和T4水稻籽粒中清蛋白、球蛋白、谷蛋白以及总蛋白质含量均增加,且T2作用效果更明显,分别较T1显著提高了0.14、0.10、0.45和0.60个百分点(2017年两品种均值);T2和T3的醇溶蛋白含量略有降低,T4则略有增加,但与T1差异均不显著;T2和T3水稻籽粒中Cu、Mn、Fe、Ca、K、Zn含量均增加,且T2增加更明显,T4水稻籽粒中各矿质元素含量变化则无明显规律。通过分析氮肥运筹下水稻籽粒植酸含量与蛋白质、8种矿质元素含量的相关性可知,植酸含量与总蛋白质、清蛋白、球蛋白、谷蛋白以及Cu、Mn、Fe、Zn、Ca、Mg、K含量均呈负相关,其中与球蛋白含量和2018年Zn和Ca含量呈显著负相关。综上所述,氮肥平衡分配能降低水稻籽粒中植酸含量,增加总蛋白质和矿质元素含量,有效改善蛋白质组分,从而提高稻米营养品质。本研究结果为寒地稻米的优质栽培的氮肥运筹方式提供了理论依据。