Assessment of soy genotype and processing method on quality of soybean tofu

Protein quality in six soybean varieties, based on subunit composition of their protein, was correlated with quality of the produced tofu. Also, protein changes due to a pilot plant processing method involving high temperature/pressure and commercial rennet as coagulant were assessed. In each soybean variety, glycinin (11S) and β-conglycinin (7S) as well as 11S/7S ratio significantly changed from beans to tofu. Between varieties, the 11S/7S protein ratio in seed indicated genotypic influence on tofu yield and gel hardness (r = 0.91 and r = 0.99, respectively; p < 0.05). Also, the 11S/7S ratio correlated with soymilk pH (r = 0.89, p < 0.05), leading to a relationship between soymilk pH with protein recovery and yield of tofu (r = 0.94 and r = 0.91, respectively; p < 0.05). The soybean β'-subunit of 7S protein negatively influenced tofu hardness (r = -0.91, p < 0.05). Seed protein composition and proportion of 7S protein subunits under the applied production method had an important role in defining tofu quality.     

Processing effect on soybean storage proteins and their relationship with tofu quality

Tofu was prepared from 13 soybean varieties according to three different methods (bench, pilot, and production methods). Different soybean varieties showed significant differences in storage protein composition (glycinin and beta-conglycinin). The beta-conglycinin (7S) and glycinin (11S) contents were 7.3-9.9 and 14.1-22.9% on the dry matter basis, respectively. The 11S/7S protein ratio varied from 1.64 to 2.51 among the varieties. Glycinin content and 11S/7S protein ratio of the 13 varieties did not change significantly from soy milk to tofu for the production and pilot methods. Soybean 11S/7S protein ratio positively correlated with the 11S/7S ratio of soy milk and tofu (0.57 < or = r < or = 0.83, p < or = 0.01). The correlation coefficient depended on the processing method. Processing method affected 7S and 11S protein contents of tofu and their contribution to tofu hardness, yield, and sensory quality. This may explain in part the contradictory findings of the relationships between storage proteins and tofu quality because processing methods differed in various studies.     

Protein recovery in soymilk and various soluble fractions as a function of genotype differences, changes during heating, and homogenization

Harovinton, a variety of tofu type soybean, and 11 derived null soybean genotypes lacking specific glycinin (11S) and beta-conglycinin (7S) protein subunits were investigated to determine whether changes in protein composition affected the protein recovery in soymilk and its soluble fractions after various centrifugation steps. As both heating and homogenization have a marked effect on the increase in protein solubility, the changes occurring during these processing steps were studied for each soybean genotype. Harovinton and 11S-null genotypes showed significantly higher protein yields than the other genotypes evaluated. Subunits of group I (A(1), A(2)) of glycinin had a negative impact on protein solubility in all treatments, but this effect was the greatest in unheated soymilk samples. Samples containing a high beta-conglycinin to glycinin ratio showed an effect of heating on the solubility of the protein, as beta-conglycinin subunits aggregate with heating. The presence of the alpha' subunit of beta-conglycinin aids in the recovery of protein in the supernatant prepared from lines containing group I (A(1,4) A(2)) glycinin. The results of this study will help determine which specific protein composition will confer an increased stability in soymilk and soymilk-derived products.     

晋豆24~(60)Coγ射线诱变后代种子贮藏蛋白的研究

选用晋豆24及其60Coγ射线辐照诱变后代为试验材料,利用SDS-PAGE聚丙烯酰胺凝胶电泳研究了诱变后代大豆贮藏蛋白亚基11S和7S的组分含量、亚基组成和各亚基的含量及其变异规律。结果表明:大豆贮藏蛋白种质资源中,同一品种的不同亚基和不同品种的同一亚基间均有较大变异;11S球蛋白与7S球蛋白呈极显著负相关,与11S/7S比值呈极显著正相关;聚类分析根据11S/7S比值将诱变后代群体分为比值较高(平均为1.70)、居中(平均为1.25)和较低(平均为0.79)3类,为诱变育种选育优质专用大豆新品种提供了依据。     

Effect of soy milk characteristics and cooking conditions on coagulant requirements for making filled tofu

The amount of coagulant added to soy milk is a critical factor for tofu-making; particularly it affects the textural properties of tofu. Earlier research indicated that the critical point of coagulant concentration (CPCC) is a characteristic parameter of soy milk and could be used as an effective indicator of optimal coagulant concentration (OCC) for making filled tofu. The objective of this study was to investigate the possible correlations between CPCC and the characteristics of soy milk made from various soybean samples and the effect of soy milk cooking and dilution conditions on CPCC. CPCC was determined by a titration method. Calcium chloride and magnesium chloride were used as coagulants. Soy milk characteristics including solid, protein, phytate, pH, titratable acidity, mineral content, and 11S/7S protein and these characteristics as affected by heating rate, heating time, and sequence of dilution and heating were studied. The results showed that the CPCC was significantly (p < 0.05) positively correlated with phytate content (grams per gram of protein), pH, and 7S protein content but negatively correlated with protein content, 11S protein content, 11S/7S ratio, titratable acidity, and original calcium content. Within the same soybean material, more proteins required more coagulant, but higher protein concentration during cooking resulted in less coagulant required by each gram of protein during coagulation. The CPCC decreased with increasing soy milk heating time or decreasing heating rate. The sequence of heating and diluting for preparing soy milk also had an effect on CPCC.     

Biochemical characterization and cellular localization of 11S type storage proteins in olive (Olea europaea L.) seeds

The composition of seed storage proteins (SSPs) in olive endosperm and cotyledon has been analyzed. Precursor forms of these proteins are made up of individual proteins, which have been purified to homogeneity and further named p1-p5 (20.5, 21.5, 25.5, 27.5, and 30 kDa, respectively). N-terminal sequences of p1 and p2 proteins displayed relevant homology to the basic subunit of the 11S family of plant SSPs (legumins). Two-dimensional polyacrylamide gel electrophoresis experiments allowed us to verify the basic character of p1 and p2 and the acidic character of p3, p4, and p5 proteins. In addition, the putative presence of highly similar isoforms or posttranslational modifications of these polypeptides was detected. As a result, a model describing the putative association of p1-p5 proteins into subunits of alpha(acidic)/beta(basic) type has been proposed. Solubility experiments have shown that the majority of these olive seed proteins from the 11S storage protein family are extracted with aqueous alcohol and only partially with water and diluted saline solutions, therefore suggesting their similarity to prolamines. Moreover, no visible differences were found in either subunit composition or 11S proteins mass among six olive cultivars examined. This result suggests that the synthesis of storage proteins is highly conserved in this plant species. By using a rabbit antiserum raised to p1 protein, the proteins have also been immunolocalized in olive seed tissues, showing that they accumulate in conspicuous protein bodies present in both the endosperm and the cotyledon.     

空间坏境对大豆主要农艺性状及蛋白品质的诱变效应

以“实践八号”育种卫星搭载的3个大豆品种中黄28、中黄29、中黄31为材料,调查了空间诱变后代SP1、SP2代的主要农艺性状分析检测SP2、SP3种子的蛋白质组分（11S/7S球蛋白）、Kunitz胰蛋白酶抑制剂（SKTI）等品质性状分析检测,以为利用空间诱变技术改良大豆品质提供理论依据。结果表明,3个大豆品种对空间环境的反应敏感性不同,中黄28和中黄29的SP1代分别获得3株和12株变异株,变异率分别为0.417 %和1.667 %,而中黄31的SP1代没有发现变异植株,且中黄28的2株（01-SP2-1、01-SP2-2）和中黄29的5株（02-SP2-2、02-SP2-5、02-SP2-6、02-SP2-7、02-SP2-8）变异株在SP2代发生性状分离;空间环境使大豆11S 蛋白亚基发生变异,SKTI基因发生突变。空间诱变既可改良农艺性状,也可改良品质性状。     

Hydroponic cultivation improves the nutritional quality of soybean and its products

Hydroponic cultivation allows the control of environmental conditions, saves irrigation water, increases productivity, and prevents plant infections. The use of this technique for large commodities such as soybean is not a relevant issue on fertile soils, but hydroponic soybean cultivation could provide proteins and oil in adverse environmental conditions. In this paper, the compositions of four cultivars of soybean seeds and their derivates, soy milk and okara, grown hydroponically were compared to that of the same cultivar obtained from soil cultivation in an open field. Besides proximal composition, the concentrations of phytic acid and isoflavones were monitored in the seeds, soy milk, and okara. Results demonstrated that, independent from the cultivar, hydroponic compared to soil cultivation promoted the accumulation of fats (from 17.37 to 21.94 g/100 g dry matter) and total dietary fiber (from 21.67 to 28.46 g/100 g dry matter) and reduced isoflavones concentration (from 17.04 to 7.66 mg/kg dry matter), whereas protein concentration was unaffected. The differences found in seed composition were confirmed in the respective okara products, but the effect of cultivation system was not significant looking at the soy milk composition. Data showed that hydroponic cultivation improved the nutritional quality of soybean seeds with regard to fats and dietary fiber. They also suggest that specific cultivars should be selected to obtain the desired nutritional features of the soybean raw material depending on its final destination.     

Two-dimensional electrophoresis and western-blotting analyses with anti Ara h 3 basic subunit IgG evidence the cross-reacting polypeptides of Arachis hypogaea, Glycine max, and Lupinus albus seed proteomes

The allergenicity of seed storage proteins, the major components of edible legume seeds, may cause serious reactions in both children and adult population. Updated methodologies for evaluation of the activity of these proteins are needed. In this paper we used two-dimensional (2D) electrophoretic techniques to investigate the immuno-cross-reactivities of anti Ara h 3 basic subunit IgG to the seed proteomes of three legume species, namely, peanut, soybean, and lupin. The seed proteins, extracted with two different procedures, were separated by 2D electrophoresis, and the electrophoretic maps were analyzed by Western blot. In peanut proteome the antibodies strongly reacted with the 23 kDa polypeptides, corresponding to Ara h 3 basic isoforms, the antigen they were raised to, and three unidentified acidic polypeptides near 45 kDa. Remarkable cross-reactivities with lupin and soybean Ara h 3 homologous polypeptides and nonrelated proteins, namely, lupin conglutin gamma and soybean Bg7S, were detected. Therefore, these proteins may be regarded as new putative allergens. The present findings show the potentiality of 2D electrophoresis in the identification of food allergens and open the way to the traceability of the new cross-reacting proteins in the food chain.     

Mungbean [Vigna radiata (L.) Wilczek] globulins: purification and characterization

Vicilin type (8S) and basic 7S globulins and legumin type (11S) globulins were isolated from mungbean [Vigna radiata (L.) Wilczek]. The native molecular weights of the different globulin types were 360000 for legumin, 200000 for vicilin, and 135000 for basic 7S. Some of the 8S globulin apparently complexed and coeluted with the 11S on gel filtration. On SDS-PAGE, 11S was composed of two bands of 40000 and 24000, 8S was composed of 60000, 48000, 32000, and 26000 bands, and basic 7S was composed of 28000 and 16000 bands. The percent composition of total globulins was estimated to be as follow: 8S, 89%; basic 7S, 3.4%; and 11S, 7.6%. The basic 7S and 11S but not the 8S globulins were found to have disulfide bonds. The presence of carbohydrates by conjugated peroxidase reaction was observed in all bands of 8S, the acidic polypeptide of basic 7S, and its complex but not in 11S. The 28000 basic 7S band and its 42000 complex and the first three major bands of 8S cross-reacted with antibodies to all types of soybean conglycinin subunits (alpha, alpha', and beta), whereas the fourth band cross-reacted only with the anti-beta subunit. None of the mungbean globulins cross-reacted with anti-soybean glycinin. Basic 7S was found to be easily extracted with 0.15 M NaCl, 11S was extracted with 0.35 M NaCl,and 8S was extracted over a wide range of NaCl concentrations. The N-terminal sequences of the different subunits/fragments of the globulins were determined and found to have strong homology with storage proteins of other legumes and crops.     

The structure of microbial communities in redoximorphic microsites of Gleysol

Microbial communities were studied in redoximorphic microsites of highly heterogeneous Gleysol at a mm scale using 16S and 18S amplicon sequencing to demonstrate if the composition of soil microbes reflects the differences in ferric and ferrous micro-sites. In both explored gley horizons with redoximorphic features (Bg2 and Cg), ferric mottles were significantly enriched with total P and Fe and depleted of O, Si, Al, K and Ca compared with the adjacent ferrous groundmass (SEM–EDS). Ferric mottles were determined as Fe oxide coatings and hypocoatings. In Bg2, both prokaryotic and micro-eukaryotic communities differed significantly between mottles and groundmass in composition of operational taxonomic units (OTUs) and in proportions of phyla, reflecting heterogeneities in the soil properties there. Mottles in Bg2 were characterized by increased proportion of Proteobacteria, decreased proportion of Acidobacteriota among prokaryotes and by dominance of a single proteobacterial OTU from Anaplasmataceae compared to all other samples. The composition of micro-eukaryotes showed an opposite trend, as micro-eukaryotes of Bg2 groundmass were unique among the other horizons, while micro-eukaryotes of Bg2 mottles had similar composition to neighbouring horizons. Microbial communities of adjacent samples were not more similar to each other than communities of randomly selected ones in Bg2 horizon. That suggests that at mm scale, the sample distance does not represent the driving factor of microbial community composition and that the adjacent samples differ rather due to physicochemical factors. The spatial organization of microbial communities revealed in Bg2 has not reappeared in similarly organized Cg horizon, probably due to other overriding factors. The differences revealed between Bg2 and Cg horizons, including granulometric composition, content of crystalline Fe, exchangeable Al, and organic carbon, as well as exposition to groundwater, were discussed as possible reasons of the distinct organization in Cg. The similarity of pro−/eukaryotic communities of adjacent and non-adjacent couples suggests no distance decay pattern at a mm scale. The agreement between patchiness in soil properties and microbial communities was revealed for the first time and confirms the importance of microscale patterns in soil.     

Thermal Behavior and Nonfreezing Water of Soybean Protein Components

Thermal denaturation and hydration of two soybean protein components were studied using differential scanning calorimetry (DSC). Results showed that temperature of denaturation (T_d) of both 11S and 7S globulins decreased sharply with an increase in water content. Enthalpy of denaturation (ΔH_d) of 11S increased with increasing water content at first, and then leveled off at high water content. However, ΔH_d of both 7S and 11S components in 7S samples first increased and then decreased at high water content. The preparation method of samples influenced the ΔH_d value significantly but had little effect on the T_d. Nonfreezing water was determined from the DSC results. It increased in both 11S and 7S as water content increased but was more abundant in 7S, probably because of different compositions and structures. Threshold value of water content for the appearance of freezing water was 0.30–0.32 h (g of water/g of protein, mass ratio) for 11S. The water absorbed by both 11S and 7S during denaturation increased quickly at low water contents and remained almost constant at high water contents. The results were attributed to different structure and conformation of proteins before and after denaturation.     

Health-promoting effects of a dietary fiber concentrate from the soybean byproduct okara in rats

Okara (a byproduct of the soy milk industry) is rich in proteins (24.5-37.5 g/100 g of dry matter (dm)), lipids (9.3-22.3 g/100 g of dm), and dietary fiber (DF) (14.5-55.4 g/100 g of dm). It also contains isoflavones (0.14 g/100 g of dm). In the present study we fed female healthy Wistar rats either a standard rat diet or a supplemented 10% DF-rich okara (DFRO) diet for 4 weeks, and then we assessed several health parameters in the serum and the cecum compartments. In comparison to the control group, rats fed DFRO showed a significant decrease in weight gain (5.00 +/- 1.22 g vs 2.00 +/- 1.46 g, P < 0.03, during week 4) and in total cholesterol (65 +/- 8 mg/dL vs 51 +/- 5 mg/dL, P < 0.05) and a significantly increased antioxidant status (36.71 +/- 15.31 micromol of Trolox equivalents (TEs)/g vs 69.75 +/- 16.11 micromol of TEs/g, P < 0.0003) and butyrogenic effect (39.37%, P < 0.003) in the cecum. In addition, a significant enhancement in the apparent absorption (41.89 +/- 1.64% vs 47.02 +/- 2.51%, P < 0.004) and in the true retention (41.62 +/- 1.60% vs 46.68 +/- 2.55%, P < 0.005) of calcium was appreciated. In summary, these findings show for the first time that a concentrate DF from a soybean byproduct protects the gut environment in terms of antioxidant status and prebiotic effect. These results may highlight the development of an innovative soybean byproduct rich in DF which could be useful as a functional ingredient with health-promoting attributes.     

Enzymatic high digestion of soybean milk residue (okara)

The objective of this study was to digest okara in high yield by food-processing enzymes. Autoclaving of okara was effective in increasing cellulase digestion for the primary cell wall, and the digestion was accelerated by the formation of single cells by stirring. Most of the residual okara after autoclaving and cellulase digestion was found to be the secondary cell walls compared with the cellulase-treated soybean single cells. The secondary cell wall was found to be composed of galacturonic acid, neutral sugars, and protein and was considered to be a complex of these compositions. Many cellulolytic and proteolytic enzymes could not digest the secondary cell wall; however, it was found that two pectinases could digest the secondary cell wall. A series of digestions resulted in yields of 83-85% from the raw okara, and the final residues were identified as oil body complexes in the soybean cells and fiber-like organ between the cells.     

Mycorrhizal fungi effects on nutrient composition and yield of soybean seeds

Nutrient composition and yield of soybean [Glycine max (L.) Merr] seeds are heritable traits affected by environmental factors. This study determined the effects of arbuscular‐mycorrhizal (AM) fungi on seed protein, lipid, and phosphorus (P) composition and yield in soybean grown under a high nitrogen (N) regime. Plants were grown in pot cultures without AM fungi in P‐fertilized (+P) or unfertilized (‐P) soil, or in ‐P soil inoculated with one of the AM fungi Glomus mosseae (Nicol. & Gerd.) Gerd, and Trappe (Gm), Glomus etunicatum Becker and Gerd.(Ge), or Gigaspora rosea Nicol. and Schenck (Gr). Seed yields of+AM plants, as a group, were halfway between those of the +P and ‐P plants. Seed size was highest in Gm plants. Differences in protein concentrations between Ge and Gr and the other treatments were highly significant. Seed P and protein concentrations were not significantly correlated (p=0.162), but a highly significant (r =‐0.949) negative correlation between seed P and lipid concentrations was observed. Phosphorus concentration was highest and that of lipids lowest in +AM plants. Seed yield and nutrient composition were independent of the intensity of root colonization. The seed protein/lipid ratio was highly correlated with seed P concentration and was significantly higher for +AM plants, as a group, than for both +P and ‐P ‐AM plants. Differences in seed dry weight, size, seed/ stem ratio, P content, and protein concentration among +AM plants showed mycorrhiza‐specific host responses. These responses suggest that AM fungi can modify soybean seed development and chemical composition.     

Effects of fly attack (Bactrocera oleae) on the phenolic profile and selected chemical parameters of olive oil

The phenolic fraction of virgin olive oil influences both its quality and oxidative stability. One of the principal threats of the quality of olive fruit is the olive fly ( Bactrocera oleae) as it alters the chemical composition. The attack of this olive pest has been studied in order to evaluate its influence on the quality of virgin olive oil (free acidity, peroxide value, fatty acid composition, water content, oxidative stability, phenols, and antioxidant power of phenolic fraction). The study was performed using several virgin olive oils obtained from olives with different degrees of fly infestation. They were acquired in different Italian industrial mills from the Abruzzo region. Qualitative and quantitative analyses of phenolic profiles were performed by capillary electrophoresis-diode array detection, and electrochemical evaluation of the antioxidant power of the phenolic fraction was also carried out. These analyses demonstrated that the degree of fly attack was positively correlated with free acidity ( r = 0.77, p < 0.05) and oxidized products ( r = 0.58, p < 0.05), and negatively related to the oxidative stability index ( r = -0.54, p < 0.05) and phenolic content ( r = -0.50, p < 0.05), mainly with secoiridoid compounds. However, it has been confirmed that the phenolic fraction of olive oil depends on several parameters and that a clear correlation does not exist between the percentages of fly attack and phenolic content.     

Quality indicators of cultured Newfoundland blue mussels (Mytilus edulis) during storage on ice: microbial growth, pH, lipid oxidation, chemical composition characteristics, and microbial fatty acid contents

To devise proper strategies to evaluate and maintain the quality of cultured blue mussels (Mytilus edulis) during storage on ice, this study examined the potential use of microbial growth, pH, lipid oxidation, and composition characteristics as quality indicators. It also evaluated the use of individual (i15:0, ai15:0, i16:0, ai16:0, i17:0, ai17:0, 17:0) and total bacterial fatty acid contents as a rapid method to examine the microbial quality of stored mussels. Linear regression analyses of the storage period of mussels vs various quality indicators indicated that the strongest correlation was between the storage period and microbial growth (r = 0.973, p < 0.0001) followed by the microbial fatty acid ai15:0 content (r = 0.903, p < 0.0001) and ash content (r = 0.819, p = 0.0002). All other correlations between the storage period and quality indicators were moderate (r = 0.500-0.700 and p < 0.05), poor (r < 0.500 and p > 0.05), or negative (r = -0.657 and p = 0.027).     

Relationships Between Soybean 11S Globulin Content and Thermal and Retrogradation Properties of Nonwaxy Maize Starch

In the present study, the relationships of soybean 11S globulin content, thermal properties, and retrogradation properties of nonwaxy maize starch in starch–globulin mixtures were investigated by differential scanning calorimetry. The onset and peak temperatures of maize starch were positively related to soybean 11S globulin content, whereas the thermal enthalpy was negatively related to it. However, the onset temperature, peak temperature, and thermal enthalpy of soybean 11S globulin were negatively related to maize starch content of mixtures. On the other hand, the onset and peak temperatures of retrograded maize starch were positively related to soybean 11S globulin content, whereas the retrogradation enthalpy was negatively related to it during storage. Therefore, adding soybean 11S globulin was an effective method to control maize starch gelatinization and retrogradation properties, which will be helpful for the food industry to produce high‐quality products based on starch and soybean protein.     

Imbibition of soybean seeds in warm water results in the release of copious amounts of Bowman-Birk protease inhibitor, a putative anticarcinogenic agent

Protease inhibitors play a protective role against pathogenic microorganisms and herbivorous insects. The two predominant protease inhibitors of soybean seeds are the Kunitz trypsin inhibitor (KTI) and Bowman-Birk protease inhibitor (BBI). In this study, we report that soybean seeds incubated in warm water release large amounts of proteins into the surrounding media. Two-dimensional gel electrophoresis analysis of the seed exudates resulted in the separation of 93 distinct protein spots out of which 90 spots were identified by LC-MS/MS. The basic 7S globulin and the BBI are the two predominant proteins found in the soybean seed exudates. In addition to 7S and 11S seed storage proteins, others known to protect the seeds against pathogens and pests including KTI, peroxidase, α-galactosidase, and endo-1.3-β-glucanase were also identified in the seed exudates. Soybean seed exudate obtained by incubating the seeds in warm water was also able to inhibit the growth of human breast cancer cell line MCF-7. Since soybean seeds release large amounts of enzymatically active BBI when immersed in warm water, our procedure could be exploited as a simplified alternative method for the preparation of BBI concentrate which is being used as a cancer chemoprotective agent.