Effects of Phytases and Dehulling Treatments on In Vitro Iron and Zinc Bioavailability in Faba Bean (Vicia faba L.) Flour and Legume Fractions

ABSTRACT: In vitro digestions were performed on faba bean flours with decreased phytate contents and on 2 dephytinized or nondephytinized faba bean fractions, a dehulled faba bean fraction, and a hull fraction with low and high fiber and tannin contents, respectively. In vitro bioavailability iron and zinc was defined as the relative amount of iron and zinc that became soluble after enzymatic treatment. Faba bean samples were sequentially digested with enzymes, including amylase, pepsin, pancreatin, and bile, under certain conditions following the enzymatic degradation procedure. Iron and zinc in vitro bioavailability of whole faba bean flours were significantly improved by phytate degradation, even if the phytate were not all degraded. Total dephytinization of dehulled faba bean led to an obvious increase in iron and zinc in vitro bioavailability, but that of hulls had no effect on either iron or zinc in vitro bioavailability. Fibers and tannins other than phytate are more important in chelating a high proportion of iron and zinc in faba bean hulls.     

Effects of phytase,cellulase and dehulling treatments on zinc in vitro solubility in faba bean (Vicia faba L.) flour and seed fractions

Simulations of gastro‐intestinal digestion was used to try to identify the nature of the complexes between antinutritional factors and zinc in faba bean fractions. In the hull fraction, the action of phytases and the simultaneous action of cellulase and phytase allowed about 7% and 35% additional zinc to be solubilised, respectively. Single enzymatic degradation of phytates from dehulled faba bean allowed solubilisation from 65% to 93% of zinc depending upon the treatment. The zinc of faba bean flour was mainly linked to phytates and the greater the degree of phytate degradation before in vitro digestion, the greater the quantity of zinc released. In dehulled faba bean zinc was almost exclusively chelated by phytates. In the hull of faba bean, the majority of zinc was chelated in complexes between phytates and fibres.     

The impact of processing on phytic acid,in vitro soluble iron and Phy/Fe molar ratio of faba bean (Vicia faba L.)

BACKGROUND: Faba bean is one of the important legumes in Asian countries. It is also a major source of micronutrients in many rural areas. Unfortunately, the bioavailability of iron from faba bean is low because it is present as an insoluble complex with food components such as phytic acid. The influence of soaking, germination and fermentation with the expectation of increasing the bioavailability of iron was investigated. RESULTS: Fermentation treatments were most effective in decreasing phytic acid (48–84%), followed by soaking at 10 °C after preheating (36–51%). Steeping faba beans for 24 h at 25 °C had the least effect on the removal of phytic acid (9–24%). With increased germination time at 30 °C, phytic acid progressively decreased from 9 to 69%. Most wet processing procedures, except soaking after wet preheating, caused losses of dry matter and iron (8–15%). In vitro iron solubility, as a percentage of total iron in soaked faba bean after dry preheating, was significantly higher than in raw faba bean (P < 0.05). Fermentation and germination did not have significant effects on the solubility of iron. CONCLUSION: The expected improvement of iron bioavailability levels due to lower phytic acid was not confirmed by increasing levels of in vitro soluble iron. Soaking, germination and fermentation can decrease phytic acid in faba bean. However, results from in vitro solubility measurement of iron showed little improvement of iron bioavailability in fermented and germinated faba beans over untreated raw faba beans (P < 0.05). It seems that componets of dietary fibre other than phytic acid are more important in binding iron. Probably, a complex association between dietary fibre and iron is the reason for the poor bioavailability of iron in faba bean. Copyright © 2009 Society of Chemical Industry     

不同发芽条件对蚕豆植酸酶活性的影响

以蚕豆为试材,研究发芽过程中植酸酶在蚕豆不同部位的变化和性质.发芽蚕豆不同部位植酸酶活性呈现先高后低的变化趋势,子叶植酸酶在发芽第6天达最高值.超声波处理对发芽蚕豆胚根、子叶和胚芽植酸酶活性有一定促进作用,其中以15min超声波处理对胚根和胚芽植酸酶活性影响最大,5min处理、10min处理和15min处理均对子叶植酸酶活性有显著影响.在发芽过程中不同光照对蚕豆进行处理,不论胚根、子叶还是胚芽,黑暗处理植酸酶活性高于光照处理.核酸抑制剂放线菌素(Act-D)和蛋白质抑制剂环已酰胺(CHM)对发芽蚕豆胚根、子叶和胚芽中的植酸酶活性有一定影响,其中以CHM的抑制效果最为明显.     

High apparent ileal digestibility of amino acids in raw and germinated faba bean (Vicia faba)‐ and chickpea (Cicer arietinum)‐based diets for rats

The effects of germinated faba bean (Vicia faba minor var Alameda)‐ and chickpea (Cicer arietinum kabuli var Athenas)‐based diets on performance, N utilisation, amino acid ileal digestibility and plasma concentrations were studied in growing male Wistar rats. Raw and germinated faba bean and chickpea seed meals were incorporated in essential amino acid‐supplemented and energy‐equalised diets as the only sources of protein. A lactalbumin‐based diet equalised in protein (100 g kg^?1) and digestible energy (15.5 kJ g^?1) with the test diets was used as control. Weight gains, gain/feed ratios, N retention and NPU values of animals fed raw or germinated legumes were lower than those of rats given the control (lactalbumin) diet, while faecal dry weights were higher. However, N retention and NPU values of rats fed germinated faba bean diets were higher and faecal dry weights lower than those of animals fed non‐germinated faba bean diets. Liver and thymus fresh and dry relative weights were lower than controls for all legume diets, whilst small intestine, caecum and colon relative weights were higher than controls for non‐germinated faba beans and chickpeas either germinated or not. Caecum and colon fresh and dry relative weights of rats fed germinated faba bean seeds were lower than those of rats fed non‐germinated faba beans and not different from controls. Except for aspartic acid and proline in rats fed raw faba bean diets, apparent ileal digestibilities of amino acids were not different or higher in rats fed legume diets as compared with controls. Except for arginine, apparent ileal amino acid digestibility was higher for germinated than for non‐germinated faba bean seeds. The amounts of glycine, arginine and ornithine in plasma significantly increased whilst those of threonine, alanine, leucine and lysine decreased with respect to controls in faba bean‐ or chickpea‐fed rats, either germinated or not. We conclude that (a) germination of faba bean seeds gave rise to a significant (P < 0.01) but limited improvement in performance, N retention and apparent ileal digestibilities of some amino acids in growing rats compared with non‐germinated seeds, while there was no detectable effect in the case of germinated chickpea seeds, and (b) the apparent ileal amino acid digestibility of essential amino acid‐supplemented diets based on faba bean or chickpea meals, either germinated or not, is similar or even higher than controls. © 2002 Society of Chemical Industry     

Impact of pre-treatment (soaking or germination) on nutrient and anti-nutrient contents,cooking time and acceptability of cooked red dry bean (Phaseolus vulgaris L.) and chickpea (Cicer arietinum L.) grown in Ethiopia

Pulses are processed in diverse ways prior to consumption. Soaking and germination are among the most common traditional, household-level food processing strategies. This study was carried out to determine the effects of soaking, germination, cooking and their combinations on the contents of selected nutrients and anti-nutrients of red dry bean and chickpea. In addition, the effects of pre-treatment on cooking time and the acceptability of dishes prepared from red dry bean and chickpea were determined. The nutrient compositions (zinc, iron and calcium) of most soaked-cooked and germinated-cooked red dry bean and chickpea samples were not significantly different than those of respective controls. However, soaking and germination pre-treatments significantly lowered the phytate and tannin contents of the red dry bean and chickpea samples, with a few exceptions, and overall, polyphenol contents were lower after soaking-cooking than after germination-cooking. Most scores for sensory attributes of bean-based and chickpea-based dishes prepared from soaked or germinated samples were not significantly different than those of the controls. For most red dry bean and chickpea samples, longer germination times yielded superior results in terms of reductions in cooking time, tannin content, and phytate:zinc and phytate:iron molar ratio.     

不同发芽条件对蚕豆植酸酶活性变化和性质的影响

以蚕豆为试材,研究了发芽过程中植酸酶在蚕豆不同部位的变化和性质。发芽蚕豆不同部位植酸酶活性呈现先高后低的变化趋势,子叶植酸酶在发芽第6天达到最高值。超声波处理对发芽蚕豆胚根和子叶植酸酶活性有一定促进作用,其中以15min超声波处理对胚根植酸酶活性影响最大,5min处理、10min处理和15min处理均对子叶植酸酶活性有显著影响。在发芽过程中不同光照对蚕豆进行处理,不论胚根、子叶还是胚芽,黑暗处理植酸酶活性高于光照处理。核酸抑制剂放线菌素(Act-D)和蛋白质抑制剂环已酰胺(CHM)对发芽蚕豆胚根、子叶和胚芽中的植酸酶活性有一定影响,其中以CHM的抑制效果最为明显。     

发芽处理对蚕豆主要营养成分与抗营养因子的影响

研究发芽处理对启豆2号蚕豆主要营养成分与抗营养因子含量的影响。对蚕豆发芽前后蛋白质、脂肪、淀粉、VC、氨基酸、植酸和单宁含量测定。结果表明：蚕豆在发芽1d时可溶性蛋白质含量显著增加,随后逐渐下降;粗脂肪和淀粉含量均呈下降趋势;发芽2d时VC含量显著增加,达到最大58.65mg/100g,随后逐渐下降,5d后VC含量仍高于未发芽的;发芽后1～2d蚕豆的氨基酸含量略有增加;发芽1d抗营养因子植酸含量下降了33.55%,随后平缓;发芽1d蚕豆的单宁含量增加到462.70mg/100g,2～3d增加比较平稳,5d急剧增加至1306.10mg/100g。可推知,将启豆2号蚕豆发芽1～2d的作为食用及加工的较佳时期。     

Effects of polyphenol oxidation on in vitro iron availability in faba bean (Vicia faba L.) flour

Tannic acid (TA) and green tea extract (GTE) were added to faba bean flour, before and after incubation with polyphenol oxidase (mushroom tyrosinase), and the effect on in vitro iron availability was investigated. The inhibitory effect of TA and GTE was dose dependent, and the in vitro iron availability decreased from 64.5% in the pure faba bean flour to 9.8% (P < 0.05) with the addition of 1 mg of TA 1 g^?1 to faba bean flour. The addition of 5 mg of GTE resulted in low in vitro iron availability as after the addition of 10 mg of TA (11.6% against 10.3% for 1 mg TA). Incubation of the polyphenols with tyrosinase before addition to the faba bean flour significantly increased the in vitro iron availability. The increase in iron availability was significant (P < 0.05) when amounts of 0.2, 0.5 or 1 mg g^?1 TA or GTE added to faba bean flour, respectively, were oxidised, even with the lowest amount of tyrosinase (150 u). Oxidation of 1 mg TA or GTE with 150 u tyrosinase increased the in vitro iron availability from 10.3% to 15.5% and from 19.2% to 26.1%, respectively. At the 300‐u level, the addition of higher amounts of enzyme (from 600 to 900 u) did not have any effect. The results from the study therefore suggest that the oxidation of polyphenols may be a promising way to increase the availability of iron in polyphenol‐containing legume foods.     

Effects of cultivar and culture conditions on γ‐aminobutyric acid accumulation in germinated fava beans (Vicia faba L.)

BACKGROUND: High levels of γ‐aminobutyric acid (GABA) accumulate in plant tissues under various stresses. GABA accumulation is also influenced by cultivar. This aim of this study was to select the most promising cultivar of fava bean for GABA accumulation and to optimise the culture conditions for GABA production in germinated fava beans by response surface methodology based on central composite design (CCD). RESULTS: GABA content and glutamate decarboxylase activity in germinated seeds of cultivar S2 were significantly higher than those in other cultivars (P < 0.05). A significant negative correlation (r = −0.765, P < 0.05) between germination percentage and 1000‐kernel weight was observed. There was a linear relationship between GABA content and sprout length (R² = 0.816). The regression model fitted to the experimental data of CCD was valid in predicting GABA production in germinated fava beans. Temperature and pH value of the culture solution had significant effects on GABA content in germinated fava beans. Under optimal culture conditions (33.6 °C, pH 3.19 and an air flow rate of 1.19 L min⁻¹), GABA content reached up to 2.41 g kg⁻¹ dry weight, about 48 times that in raw seeds. CONCLUSION: Germinated fava bean is a good resource of GABA‐rich food. Both cultivar and culture conditions have significant effects on GABA production. Copyright © 2009 Society of Chemical Industry     

Effect of Soaking and Germination on Polyphenol Content and Polyphenol Oxidase Activity of Mung Bean (Phaseolus Aureus L.) Cultivars Differing in Seed Color

Five mung bean cultivars varying in seed coat color were analyzed for polyphenol content. The effect of soaking and germination on the polyphenol content and polyphenol oxidase activity was investigated. The level of total polyphenols was found to be in the range of 310–340 mg/100 g in whole seeds. Soaking reduced the total polyphenols, whereas germination for 48 h increased total polyphenol content by 41–76%. The hydrolysable tannins, condensed tannins, and hydrolysable tannins/condensed tannins index increased with an increase in period of germination. The total and percent condensed tannins content slightly decreased while hydrolysable tannins increased during germination. Maximum polyphenol oxidase activity of 102–108 units was observed in 24 h germinated seeds. There was no significant correlation found between the polyphenolic content and polyphenol oxidase activity in germinating mung bean seeds. These findings demonstrated that the mung beans are fair sources of polyphenols, thus having great potential as a source of natural antioxidants.     

红豆皮多酚提取工艺优化及抗氧化活性分析

采用超声辅助提取工艺提取红豆皮中的多酚化合物,选取乙醇体积分数、超声功率、超声时间、液料比进行单因素试验,在此基础上,采用响应面法优化工艺参数。结果表明,红豆皮多酚超声辅助提取最佳工艺参数:乙醇体积分数60%,超声功率360 W,超声时间30 min,液料比30∶1(mL/g),在此条件下的多酚提取量为(145.28±2.21)mg/g。采用高效液相色谱法测定红豆皮多酚中13种单体酚:没食子酸、芦丁、儿茶素、绿原酸、原花青素B2、表儿茶素、阿魏酸、异牡荆素、牡荆素、异鼠李素、金丝桃苷、山奈酚、槲皮素。抗氧化试验结果表明红豆皮多酚的1,1-二苯基-2-三硝基苯肼自由基、超氧阴离子自由基清除能力略低于VC,而2,2′-联氨-双-(3-乙基苯并噻唑啉-6-磺酸)二胺盐自由基(ABTS+·)、羟自由基清除能力高于VC,说明从红豆皮中提取的多酚化合物具有较强的抗氧化活性,是一种极具开发潜力的天然抗氧化剂。     

Purification of diamine oxidase and its properties in germinated fava bean (Vicia faba L.)

Background: γ‐Aminobutyric acid (GABA) is a non‐protein amino acid with bioactive functions for human health. Diamine oxidase (DAO, EC 1.4.3.6) is one of the key enzymes for GABA formation. In the present study, this enzyme was purified from 5 day germinated fava bean and its properties were investigated in vitro. Results: The molecular mass of the enzyme estimated by Sephadex G‐100 gel filtration was 121 kDa. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) displayed a single band at a molecular mass of 52 kDa. The enzyme had optimal activity at 40 °C and retained its activity after being incubated at 30 °C for 30 min. It showed higher activity at pH 6.5 than at other pH values. The enzyme was significantly inhibited by Mg²⁺, Cu²⁺, Fe³⁺, aminoguanidine, ethylene glycol tetraacetic acid (EGTA), ethylene diamine tetraacetic acid disodium salt (EDTA‐Na₂), L ‐cysteine and β‐mercaptoethanol. The K_m value of DAO was 0.23 mmol L^?1 for putrescine and 0.96 mmol L^?1 for spermidine. However, the enzyme did not degrade spermine. Conclusion: DAO from germinated fava bean was purified. The optimal reaction temperature and pH of the enzyme were mild. The enzyme had higher affinity to putrescine than to spermidine and spermine. Copyright © 2011 Society of Chemical Industry     

Effect of soaking, germination, autoclaving and cooking on chemical and biological value of guar compared with faba bean

Raw, soaked, germinated, autoclaved and cooked both of guar and faba bean were chemically and biologically evaluated. Crude protein, crude fiber and ash contents of guar and faba bean decreased as a result of all treatments with the exception of protein content in both germinated guar and faba bean. A slight decrease in lipid content was noticed during germination, while there were no changes in lipids after soaking, autoclaving and cooking. Protein content of guar was higher than of faba bean, and showed higher values for all amino acids than faba bean protein except for lysine and tryptophan. Germination caused a slight increase in total essential amino acids, while soaking caused a slight decrease in both of guar and faba bean. There was a slight decrease in the mineral contents by the heat processes with the exception of Mg and Mn for guar and Na, Mn for faba bean due to the cooking process. Germinated guar and faba bean showed noticeable decrease in Na, K, Cu, Mn and Mg and a slight increase in Fe and Zn than the raw materials. Germination was more effective in reducing phytic acid than heat treatment, and therefore it improves the nutritional quality of both guar and faba bean. Protein efficiency ratio (PER), true digestibility (DT) and biological value (BV) were estimated in all treatments under study using rat bioassay procedures. Data revealed pronounced improvements of the nutritive value as a result of all processes under study especially germination being the most effective.     

Effect of Germination on Chemical Composition,Biochemical Constituents and Antinutritional Factors of Soya Bean (Glycine max) Seeds

The published scientific data concerning the effects of germination on chemical composition, biochemical constituents and anti-nutritional factors of soya bean are reviewed. The amino acid profile did not change to a great extent; only a noticeable increase in aspartic acid was observed whereas there was a gradual decrease in the available lysine level and lipid content as germination progressed. Both the total protein content and the nonprotein nitrogen increased after 5 days of germination. Dietary fibres are partially degraded in germinated seeds. Germinated soya bean is an excellent source of ascorbic acid and riboflavin. Niacin contents increased distinctly after germination. Germination induced a reduction in lipase inhibitor activity. The galactosyl oligosaccharides drastically decreased in germinated seeds. After 4 days of germination, the activity of certain lectins decreased to 4% of that of ungerminated soya beans. The phytic acid in the seeds was degraded by the phytase activated during germination, thus increasing the availability of the minerals present in the germinated seeds. Germination can degrade both Kunitz soya bean trypsin inhibitor and the major Bowman–Birk soya bean trypsin inhibitor; the degradation is enhanced, if germination process lasts more than 4 days. © 1997 SCI.     

Changes in quantities of inositol phosphates during maturation and germination of legume seeds

 The changes in the quantities of inositol phosphates during the maturation and germination of pea, faba bean and lupin seeds were determined in two consecutive (1993 and 1994) years of differing weather conditions. Irrespective of the year, all seeds accumulated predominantly inositol hexaphosphate (IP₆). The weather conditions influenced the accumulation of inositol phosphates in maturing seeds, but they did not influence the total content. Gradual degradation of inositol phosphates occurred during seed germination. After 8 days of germination, IP₆ was degraded by some 80% in peas, 78% in faba beans and 42% in lupin seeds. The enzymic hydrolysis of higher forms of inositol phosphates (IP₆ and IP₅) in germinating seeds was assumed to yield inositol tetraphosphate (IP₄) and inositol triphosphate (IP₃), because the quantities of these compounds increased during seed germination. Received: 1 August 1997 / Revised version: 13 October 1997     

蚕豆皮原花青素提取工艺优化

以蚕豆皮为材料,探讨微波预处理-乙醇提取法提取原花青素的工艺条件。分别考察预处理工艺中微波功率、微波时间、加水量,提取过程中的料液比、提取温度、提取时间对原花青素提取率的影响。结果表明：微波预处理-乙醇提取最佳工艺为微波功率140W、微波处理时间50s、加水量2mL/g、液料比21:1(mL/g)、温度55℃、70%乙醇溶液提取72min,原花青素提取率为94.53%。     

Effect of phytase treatment on iron bioavailability in faba bean (Vicia faba L.) flour

The effect of dephytinisation, using an exogenous phytase under optimal conditions (pH 5.5, 37 °C), and subsequent removal of the soaking solution after processing, on the bioavailability of iron from faba bean (Vicia faba L.) flour was studied. Soaking of the faba bean flour led to a considerable reduction in the content of iron (39%), whereas a lower reduction in iron content (10%), was obtained after additional treatment with phytase, than in the soaked faba bean flour. The digestive utilisation of iron from the raw and soaked faba bean flours by growing rats was negligible, but increased significantly as a result of phytase treatment. The low iron absorption obtained for the former two treatments, during an experimental period of 10 days, was not reflected in any of the haematological indices (red blood count, haemoglobin, haematocrit) or tissues (femur, heart, kidney) studied, with the exception of the sternum. The latter appears to be a useful indicator of iron bioavailability.     

In vitro starch digestibility of fresh and sun‐dried faba beans (Vicia faba L.)

Fresh and sun‐dried faba beans (Vicia faba L.) were cooked, stored for various times at 4 °C and analysed for available starch (AS), resistant starch (RS) and fibre‐associated resistant starch (FARS) contents as well as α‐amylolysis. Fresh beans required a shorter cooking time (25 min) than dried beans (158 min). Cooked fresh faba beans had a higher AS content than cooked dried faba beans. The AS content in both decreased during cold storage, with fresh beans showing a smaller decrease than dried beans with increasing storage time. Cooked fresh faba beans also had a higher total RS content than cooked dried faba beans, although a greater increase in RS content was recorded in the latter upon storage. Starch retrogradation was more prominent in cooked dried faba beans than in cooked fresh faba beans, as indicated by the consistently higher FARS content. The α‐amylolysis rate decreased with increasing storage time, i.e. long‐stored (72 h) cooked faba beans exhibited slower starch digestion, and differences were recorded between fresh and dried beans. The predicted glycaemic index ranged between 60.9 and 58.0% for cooked fresh faba beans and between 57.9 and 55.8% for cooked dried faba beans, which is suggestive of slow glucose release from starch in faba beans. Copyright © 2007 Society of Chemical Industry     

右旋糖酐/蚕豆蛋白复合凝胶的流变特性

为研究乳酸菌右旋糖酐对蚕豆蛋白食品相关性质的影响，采用哈克流变仪和质构仪等测定了添加不同浓度右旋糖酐时GDL诱导的酸致蚕豆蛋白复合凝胶质构和流变特性等指标。结果表明：添加右旋糖酐能显著增加蚕豆蛋白凝胶保水性，空白组蚕豆蛋白凝胶保水性为60.38%，1%右旋糖酐与蚕豆蛋白形成的复合凝胶保水性为70.08%（p<0.05）；右旋糖酐浓度在0.25%~1%之间，对复合蛋白凝胶色泽有一定影响；0.5%右旋糖酐与蚕豆蛋白形成的复合凝胶强度为0.27 N，与空白组蚕豆蛋白凝胶（0.37 N）差异显著，可软化含高蚕豆蛋白食品质构特性；右旋糖酐/蚕豆蛋白复合凝胶流动曲线符合carreau模型（R2>0.99），具有假塑性流体的特性；应变扫描的弹性模量G''均高于黏性模量G''，说明右旋糖酐/蚕豆蛋白复合凝胶的弹性占主导；频率扫描结果显示添加右旋糖酐可软化蚕豆蛋白凝胶，使凝胶G''、G''降低，更易于加工。在蚕豆食品中添加右旋糖酐可改善蚕豆蛋白的质构和流变特性，为拓展其应用范围提供参考。