Allergenicity of Maillard reaction products from peanut proteins

It is known that peanut allergy is caused by peanut proteins. However, little is known about the impact of roasting on the allergenicity of peanuts. During roasting, proteins react with sugars to form Maillard reaction products, which could affect allergenicity. To determine if the Maillard reaction could convert a nonallergenic peanut protein into a potentially allergenic product, nonallergenic lectin was reacted with glucose or fructose at 50 degrees C for 28 days. Browning products from heat-treated peanuts were also examined. The products were analyzed in immunoblot and competitive assays, using a pooled serum (i.e., IgE antibodies) from patients with peanut anaphylaxis. Results showed that the products were recognized by IgE and had an inhibitory effect on IgE binding to a peanut allergen. Thus, the findings suggest that these Maillard reaction products are potentially allergenic and indicate the need to verify whether the Maillard reaction products formed in peanuts during roasting increase their allergenicity.     

Transglutaminase polymerization of peanut proteins

Transglutaminase promotes protein cross-linking reactions through an acyl transferase mechanism involving protein-bound glutaminyl residues and primary amines including the epsilon-amino group of lysine residues in soy, myosin, gluten, oat globulin, casein, and whey. Herein, we present a first report of exogenous transglutaminase catalysis of several peanut protein fractions, including purified Ara h 1. In most cases, SDS-PAGE banding patterns revealed the formation of high molecular weight polymers while catalysis of Ara h 1 resulted in distinct dimer formation. Cross-linking effects were accomplished in the presence and absence of the reducing reagent, dithiothreitol. Ortho-phthaldialdehyde assays, used to quantify the degree of polymerization, indicated approximately 21% and approximately 30% coupling over a similar time interval, using either cold hexane extracted peanut protein fractions or lightly roasted flour dispersions, respectively. Rheological measurements established that transglutaminase-modified peanut extracts exhibited lowered viscosity readings compared to nontreated dispersions. Peanut protein polymers and glycoprotein conjugates, created by covalent linkage between protein substrates and monosaccharide amino sugars, exhibited similar IgE binding activity, compared to control solutions. These results suggested that potential allergic responses were not enhanced after enzymatic modification. Ultimately, these approaches may provide novel peanut-based food ingredients with unique functional characteristics for expanded applications within the world marketplace.     

Stepwise extraction of proteins and carbohydrates from soybean seed

The stepwise hot water extraction of soybeans, which were extractions in a series of procedures of whole soybean seeds, dehulled and sliced ones, and pressed ones carried out by autoclaving, was investigated to study the localization in the seed and their characteristics. The characteristics of each extraction were studied by HPLC, SDS-PAGE, components analysis, microscopic observation, and effect for some enzymes. Carbohydrates were easier to extract than protein. In the extractions, the ratio of uronic acid per total sugar was constantly about 0.3. A comparison of these extracts, soybean milk, extraction from defatted soybean meal, and soybean milk residues was also carried out, and the characteristics and the localization were investigated. Mid-sized proteins in soybean milk were easy to extract. However, hardly any high molecular weight proteins or high molecular weight carbohydrates were extracted. The proteins and carbohydrates were considered to be localized in the middle lamella and in the protein and/or oil bodies of the cell, and the proteins and carbohydrates were gradually extracted through seed and cell breaking. Gelation was observed only in the boiled extracts from whole seeds. Pepsin and trypsin digests of the high molecular weight protein had inhibitory activity against the angiotensin I converting enzyme.     

镉汞胁迫对花生种子的毒害效应

采用室内水培法,分别研究了镉(Cd2+)、汞(Hg2+)单一胁迫对花生种子萌发与幼苗的主要生理生化特性的影响。结果表明,当Hg2+浓度达到5 mg/L、Cd2+浓度达到或大于20 mg/L时出苗率下降;当Hg2+浓度达到25 mg/L、Cd2+浓度大于10 mg/L时不仅显著抑制生长,而且根系变黑腐烂;不同浓度Hg2+与叶绿素a的含量呈显著负相关,而与叶绿素b的含量呈显著正相关;不同浓度Hg2+与花生叶片过氧化氢酶活性呈极显著正相关。过氧化氢酶活性和叶绿素含量都与Cd2+浓度呈极显著负相关。     

Characterization of the proteins from Vigna unguiculata seeds

The proteins from Vigna unguiculata (L.) Walp. (cowpea) seeds were investigated. Globulins constitute over 51% of the total seed protein, with albumins composing approximately 45%. The globulins may be fractionated by native electrophoresis or anion exchange chromatography into three main components, which were termed (in decreasing order of anodic mobility) alpha-vignin, beta-vignin, and gamma-vignin. alpha-Vignin, with a sedimentation coefficient of 16.5S, is a major, nonglycosylated globulin, composed of a major 80 kDa subunit, which upon reduction, produces two polypeptides (20 and 60 kDa). beta-Vignin, with a sedimentation coefficient of 13S, is a major, glycosylated globulin, composed of two main polypeptides (55 and 60 kDa) with no disulfide bonds. Finally, gamma-vignin, a minor globulin, is composed by one main type of subunit (22 kDa), which upon reduction, is converted into a single, apparently heavier polypeptide chain (30 kDa) due to the presence of an internal disulfide bond. Immunological analyses revealed structural homology between beta-vignin and beta-conglutin (the vicilin from Lupinus seeds) but not between alpha- or gamma-vignins and their Lupinus counterparts. Haemagglutination activity toward trypsinized rabbit erythrocytes was found exclusively in the albumin fraction and was strongly inhibited by N-acetylglucosamine or chitin.     

Characterization of polymeric proteins from vitreous and floury sorghum endosperm

Differences in protein content and composition between vitreous and floury endosperm were investigated using a number of different techniques. Differences in protein cross-linking between vitreous and floury endosperm were investigated using differential solubility, size exclusion chromatography (SEC), and analysis of sulfhydryl content and composition. Vitreous endosperm was found to have higher levels of total protein and kafirins, but floury endosperm had a higher proportion of gamma-kafirins than the vitreous. Floury endosperm was found to have higher levels of SDS-soluble proteins than SDS-insoluble proteins extracted using sonication than vitreous endosperm. Conversely, vitreous endosperm had a greater proportion of the insoluble proteins. SEC analysis of the polymeric proteins revealed that the insoluble proteins had more polymeric proteins than did the soluble proteins, indicating greater cross-linking and a larger Mw distribution. Vitreous endosperm was also found to have a greater percentage (i.e., a higher ratio of disulfide to total sulfhydryls) of disulfide bonds than floury endosperm. These results show that the proteins in vitreous endosperm have a higher degree of cross-linking and a greater Mw distribution than those found in floury endosperm.     

花生种子颗粒离散元仿真参数标定与试验

由于花生排种装置在优化设计过程中缺乏准确的仿真模型参数,从而造成仿真与理论计算结果存在较大误差,一定程度上制约了花生排种装置的发展。该研究系统测定了花生种子的三轴尺寸、颗粒密度、弹性模量、泊松比等本征参数及其静摩擦因数、滚动摩擦因数、恢复系数。通过开展花生种子颗粒堆积试验,标定得到花生种间静摩擦因数为0.213,种间滚动摩擦因数为0.035。为检验标定参数的可靠性,开展了花生堆积角仿真与物理试验对比,结果表明花生物理堆积角和仿真堆积角相对误差为0.22%。通过开展机械式花生精量排种器的仿真与台架排种性能的对比试验,得到排种性能中漏播指数、重播指数相对误差分别为8.24%、5.12%,结果表明花生标定参数具有可靠性。该研究结果可为排种装置的优化设计与仿真研究提供理论参考。     

土壤种子库研究的热点问题及发展趋向

土壤种子库是指存在于土壤上层凋落物和土壤中全部存活种子的总和。土壤种子库是种群定居、生存、繁衍和扩散的基础,也是植被潜在更新能力的重要组成部分,在植被的更新、演替和恢复过程中起着重要的作用。本文总结提出了土壤种子库研究的几个热点问题以及发展趋向,包括土壤种子库的研究方法,入侵植物的土壤种子库,种子雨、种子库与地上植被的关系,种子库在退化生态系统植被恢复和生物多样性保护中的作用,土壤侵蚀对土壤种子库的影响,全球变化对土壤种子库的影响。同时对这些领域的研究进展进行了综述,旨在为进一步推动今后土壤种子库的研究工作提供参考。     

Identification of an antioxidant,ethyl protocatechuate,in peanut seed testa

The antioxidant activity and identification of the antioxidant component of peanut seed testa were investigated. The antioxidant activity of peanut seed testa was studied in the linoleic acid model system by using the ferric thiocyanate method. Among the five organic solvent extracts, the ethanolic extracts of peanut seed testa (EEPST) produced higher yields and stronger antioxidant activity than other organic solvent extracts. EEPST was separated into 17 fractions on silica gel column chromatography. Fraction 17, which showed the largest yield and significant antioxidant activity, was separated by thin-layer chromatography. Four major antioxidative subfractions were present. Subfraction 17-2 was found to be effective in preventing oxidation of linoleic acid. This subfraction was further fractionated and isolated and characterized by UV, MS, IR, and (1)H NMR techniques. The active compound was identified as ethyl protocatechuate (3,4-dihydroxybenzoic acid ethyl ester).     

Influence of thermal processing on the allergenicity of peanut proteins

Peanuts are one of the most common and severe food allergens. Nevertheless, the occurrence of peanut allergy varies between countries and depends on both the exposure and the way peanuts are consumed. Processing is known to influence the allergenicity of peanut proteins. The aim of this study was to assess the effect of thermal processing on the IgE-binding capacity of whole peanut protein extracts and of the major peanut allergens Ara h 1 and Ara h 2. Whole proteins, Ara h 1, and Ara h 2 were extracted and purified from raw, roasted and boiled peanuts using selective precipitation and multiple chromatographic steps, and were then characterized by electrophoresis and mass spectrometry. The immunoreactivity of whole peanut extracts and purified proteins was analyzed by the enzyme allergosorbent test (EAST) and EAST inhibition using the sera of 37 peanut-allergic patients. The composition of the whole protein extracts was modified after heat processing, especially after boiling. The electrophoretic pattern showed protein bands of low molecular weight that were less marked in boiled than in raw and roasted peanuts. The same low-molecular-weight proteins were found in the cooking water of peanuts. Whole peanut protein extracts obtained after the different processes were all recognized by the IgE of the 37 patients. The IgE-binding capacity of the whole peanut protein extracts prepared from boiled peanuts was 2-fold lower than that of the extracts prepared from raw and roasted peanuts. No significant difference was observed between protein extracts from raw and roasted peanuts. It is noteworthy that the proteins present in the cooking water were also recognized by the IgE of peanut-allergic patients. IgE immunoreactivity of purified Ara h 1 and Ara h 2 prepared from roasted peanuts was higher than that of their counterparts prepared from raw and boiled peanuts. The IgE-binding capacity of purified Ara h 1 and Ara h 2 was altered by heat treatment and in particular was increased by roasting. However, no significant difference in IgE immunoreactivity was observed between whole protein extracts from raw and roasted peanuts. The decrease in allergenicity of boiled peanuts results mainly from a transfer of low-molecular-weight allergens into the water during cooking.     

受钙影响的花生生殖生长及种子素质研究

采用砂培试验结合电子探针等分析方法研究受钙影响的花生生殖生长及种子素质。结果表明 ,低钙导致花生花粉外观变形 ,花粉壁松弛 ,淀粉粒小而稀少 ;电子探针下花生壁K峰升高 ,Ca、P、S和Si峰降低 ,从而影响花粉活力。在一定供钙范围 ,花生总花数、可育花数及产量随供钙增加而增加 ,影响总花数临界供钙水平为 0.6cmol(+) /kg ;影响可育花数及产量的临界供钙水平平均为 1.2 (+) /kg。花生种子钙含量与供钙水平呈显著直线相关 ,其直接影响种子发芽率和出苗率。 95%以上的花生发芽率和出苗率分别要求种子钙含量不低于 235和 278mg/kg     

钼酸钠浸种对花生种子萌芽及幼苗生长的影响

本文研究不同浓度钼酸钠浸种对花生种子萌发和幼苗生长的影响。试验结果表明,一定浓度的钼酸钠溶液浸种,对花生种子萌发具有促进作用,可以提高花生种子的发芽率,促进种子胚根的生长。在花生幼苗生长阶段,它能增加叶片叶绿素含量和可溶性蛋白质含量,提高了过氧化氢酶活性和硝酸还原酶活性。实验结果显示,钼酸钠溶液浸种是促进花生种子萌发和幼苗植株生长的重要措施。     

种下分层施用有机肥和化肥对花生产质量的影响

采用花生营养分层调控-起垄-播种一体机,通过研究花生种下分层施用有机肥与化肥对花生生长发育、养分积累、产量和品质、土壤理化性状的影响,旨在为砂姜黑土区夏花生微域土壤调理、高效施肥提供科学依据。结果表明,与有机肥和化肥撒施相比,种下施肥能改善花生农艺性状、提高经济性状,改善土壤理化性状,增加花生养分积累量,油酸含量平均增加1.94个百分点,产量平均增加554.0 kg/hm²,增产11.2%,其中,花生种下5和8 cm分层施用有机肥600 kg/hm²和化肥600 kg/hm²产量最高,为5759.4 kg/hm²。花生种下5和8 cm分层施用有机肥和化肥的产量比种下5 cm同层施用有机肥和化肥的产量平均增加175.7 kg/hm²,平均增产3.3%,油酸含量平均增加0.89个百分点。化肥减施25%时,花生种下施用有机肥和化肥比有机肥和化肥撒施产量平均增加258.1～419.4 kg/hm²,平均增产5.2%～8.5%,油酸含量平均增加0.75～1.71个百...     

Characterization and quantification of proteins in lecithins

Several methods for extraction and quantification of proteins from lecithins were compared. Extraction with hexane-2-propanol-water followed by amino acid analysis is the most suitable method for isolation and quantification of proteins from lecithins. The detection limit of the method is 15 mg protein/kg lecithin, and the quantification limit is 50 mg protein/kg. The relative repeatability limits for samples containing 0-500 and 500-5000 mg protein/kg sample were 12.6 and 7.5%, respectively. The protein recovery ranged between 101 and 123%. The protein content has been determined in different kinds of lecithins. The results were as follows: standard soy lecithins (between 232 and 1338 mg/kg), deoiled soy lecithin (342 mg/kg), phosphatydylcholine-enriched soy lecithins (not detectable and 163 mg/kg), sunflower lecithins (892 and 414 mg/kg), and egg lecithin (50 mg/kg). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein patterns of the standard soy and sunflower lecithins are very similar to those of soy flour. The protein profile of the egg lecithin shows several bands with a broad range of molecular masses. The molecular masses of the main proteins of soy lecithins and soy flour have been determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and ranged from 10.5 to 52.2 kDa. Most of the major proteins from soy and sunflower lecithins identified by MALDI-MS and electrospray tandem MS belong to the 11S globulin fraction, which is one of the main fractions of soy and sunflower seeds. In addition, the seed maturation protein P34 from the 7S globulin fraction of soy proteins has also been identified in soy lecithins. This protein has been reported as the most allergenic protein in soybean.     

Comparative study of two methods for extraction of aflatoxin from peanut meal and peanut butter

The difference between the CB and Best Foods methods in extracting aflatoxins from peanut products has been studied. The CB method yields 60, 121, 35, and 22% higher results for aflatoxins B1, B2, G1, and G2, respectively for 4 samples of peanut meal and 6 samples of peanut butter studied. Both reverse phase liquid chromatography and thin layer chromatography were used to quantitate the extracted aflatoxins.     

Interactions of grape seed tannins with salivary proteins

To evaluate the amount and type of condensed tannins binding salivary proteins, which are supposed to be involved in astringent sensation, model systems allowing further analyses of proteins and condensed tannins were developed. The precipitates formed after addition of grape seed tannins to salivary proteins indicate that a binding interaction occurs. Dissociation of insoluble complexes was achieved by sodium dodecyl sulfate treatment. Thiolysis reaction allowed the quantification and characterization of proanthocyanidins on both the resulting pellet and the supernatant. Binding proteins were investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The higher polymerized tannins predominantly precipitated together with the salivary proteins. The condensed tannins remaining in solution were low molecular weight polymers.     

Calcium- and magnesium-dependent aggregation of legume seed storage proteins.

The solubility characteristics and sedimentation behavior of total or individual globulins from legume seeds [Lupinus albus L., Pisum sativum L., and Glycine max (L.) Merr.] were investigated. The typical insolubility of globulins detected during their extraction seems to be due to the presence of a low molecular weight factor(s) in the seed extract. The solubility of the purified globulins decreases with increasing concentrations of calcium and/or magnesium, but not of other cations, showing minimum values at concentrations that vary with the particular globulin considered. Ultracentrifugation analyses revealed that the Ca(2+)- and/or Mg(2+)-induced insolubilization of the globulins involves the formation of high-order aggregates of molecules of the same or of different globulins. These macromolecular structures are dissociated under conditions of high ionic strength, suggesting the involvement of electrostatic interactions in the aggregation process. The degree of association relies heavily on the amount of Ca(2+) and/or Mg(2+) available, on the presence of chelating agents for these divalent cations, and on the ionic strength of the surrounding medium. The possible physiological significance of the findings is discussed.     

Identification of methanol-soluble compounds in sesame and evaluation of antioxidant potential of its lignans

The methanol extract of sesame (Sesamum indicum) seeds was fractionated and purified with the assistance of conventional column chromatography to afford 29 compounds including seven furofuran lignans. Among these isolates, (+)-samin (1) was obtained from the natural source for the first time. In addition, (-)-asarinin (30) and sesamol (31) were generated by oxidative derivation from (+)-sesamolin (2) and (+)-sesamin (3), two abundant lignans found in sesame seeds. To evaluate their in vitro antioxidant potential, the seven isolated lignans (1-7) and the two derivatives (30 and 31) were examined for the scavenging activities on DPPH free radicals and superoxide anions. Moreover, the capability of chelating ferrous ions and reducing power of these sesame lignans were also measured. The results suggest that, besides the well-known sesamolin and sesamin, the minor sesame lignans (+)-(7S,8'R,8R)-acuminatolide (5), (-)-piperitol (6), and (+)-pinoresinol (7) are also adequate active ingredients and may be potential sources for nutritional and pharmacological utilization.