小麦谷蛋白亚基的组成及含量与谷蛋白聚合体大分子的关系及其对面包烘焙品质的影响

含有高分子量谷蛋白亚基(HMW-GS)5 10的生物型小麦与2 12的生物型小麦相比，前者谷蛋白具有更大的分子量分布。高低分子量谷蛋白亚基的比例对于谷蛋白聚合体分子量的大小起着重要的作用，谷蛋白聚合体的体积越大，含有的高低分子量谷蛋白亚基的比例越高。SDS非可溶性谷蛋白含有较高比例的高低分子量谷蛋白亚基，并且其分子量要比可溶性谷蛋白聚合体的大。谷蛋白聚合体分子量分布的差异是不同小麦品种面包烘焙品质存在差异的重要因素。     

High‐molecular‐weight glutenin subunit composition of Pakistani hard white spring wheats grown at three locations for 2 years and its relationship with end‐use quality characteristics

Eleven Pakistani hard white spring wheat cultivars, along with one durum wheat and two hard white American‐grown wheat cultivars, were evaluated for their high‐molecular‐weight (HMW) glutenin subunit composition via sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE). The relationships among different quality characteristics and between these characteristics and HMW glutenin subunits were computed. Three to six HMW glutenin subunits were observed in Pakistani bread wheat cultivars. The presence of HMW glutenin subunits was not affected by growth locations or crop years. However, variations in intensities were observed. Correlations were noticed between certain HMW glutenin subunits and some quality attributes, such as protein, farinograph dough development time, farinograph water absorption, loaf volume and mixograph peak height. The presence of HMW glutenin subunit 20 in the older wheat cultivars C591 and C273, known for excellent chapati quality, indicated a possible relationship between this band and chapati quality. This observation will need to be confirmed by testing a larger number of wheat samples known to have characteristics for both good and poor chapati quality. © 2000 Society of Chemical Industry     

Role of gluten and its components in influencing durum wheat dough properties and spaghetti cooking quality

Protein is an important component of grain which affects the technological properties of durum wheat. It is known that the amount and composition of protein can influence dough rheology and pasta quality but the influence of the major classes of protein is not well documented. The influence of the various gluten components on dough and pasta properties was investigated. The protein composition of durum semolina was altered by either adding gluten fractions to a base semolina or preparing reconstituted flours with varying protein composition. The effects on semolina dough rheology and spaghetti texture were measured. Published methods to isolate relatively pure quantities (gram amounts) of glutenin, gliadin, high molecular and low molecular weight glutenin subunits were evaluated and modified procedures were adopted. Reconstituted flours with additional glutenin increased dough strength while additional gliadin and LMW‐GS decreased strength. These changes did not impact on spaghetti texture. Results from using the addition of protein fractions to a base semolina showed that gluten and glutenin addition increased the dough strength of a weak base semolina while gliadin addition weakened the base dough further. Addition of HMW‐GS greatly increased dough strength of the base while addition of LMW‐GS greatly reduced dough strength. Again, these affects were not translated into firmer pasta. Copyright © 2007 Society of Chemical Industry     

Relationship between gluten strength and pasta firmness in Indian durum wheats

Selected Indian durum wheats including five newly released varieties and seven landraces were studied for their grain quality, gluten strength (sodium dodecyl sulphate‐sedimentation and mixograph) and pasta making properties. Landraces were found to have good grain size, protein content, and gluten strength but they had lower test weight and semolina yield than released varieties. As protein composition affects gluten strength, landraces having combination of low molecular weight (LMW‐GS) with 5, 12, 15 and 19 linked to Gli‐B1 43.5 and high molecular weight (HMW‐GS) 2*, 14 + 15 provide a dough strength comparable with the 7 + 8 and LMW‐GS 2, 4, 6, 12, 15 and 19 (caa) allelic pattern, typically associated with good gluten strength. Landraces have rare combinations of glutenins and gliadins, which are not seen in commercial Indian durums, and in some cases, these rare alleles seem to favour good gluten strength and pasta firmness. Introduction of these alleles through breeding should improve the gluten strength and pasta making properties of Indian durum cultivars.     

Relationships between storage protein composition,protein content,growing season and flour quality of bread wheat

The storage protein composition from the Glu‐1, Glu‐3 and Gli‐1 loci encoding high and low molecular weight glutenin subunits (HMW‐GS and LMW‐GS) and gliadins, respectively, was determined on 30 wheat (T aestivum L) genotypes from three growing seasons. The gliadins and the LMW‐GS were identified as gliadin/LMW‐GS pairs. All samples were analysed by two one‐dimensional electrophoretic techniques, and selected samples were also subjected to two‐dimensional electrophoretic separation. Different statistical/data‐analytical techniques were evaluated in the study of how the presence or absence of the protein alleles, the protein content and the growing seasons are related to flour quality. The year of growth had a large impact on mixograph peak time. When predicting mixograph peak time from the presence or absence of significant proteins and the year of growth, 70% of the variability in mixograph peak time could be explained, whereas only 49% of the variability could be explained when the year of growth was deleted from the model. Protein had no effect on mixograph peak time as expected, and the well‐known positive effect of HMW‐GS 5 + 10, and the negative effects of 2 + 12 and 6 + 8 was observed. Furthermore, some of the gliadin/LMW‐GS combinations influenced mixograph peak time significantly. The gliadin/LMW‐GS at the combined Gli‐A1, Glu‐A3 loci b; ?? was positively related to mixograph peak time, whereas ??; ?? and a;a was negatively related. Although the LMW‐GS component ?? of the alleles b; ?? and ??; ?? alleles appear similar on one‐dimensional gels, two‐dimensional separation of selected samples may suggest that the ?? components in these alleles are different proteins. Cross‐validated partial least squares regression combined with empirical uncertainty estimates (jack‐knifing) of the parameters estimated in the model, gave similar results to ANOVA in identifying quality related protein alleles. The applicability of the multivariate approach in proteomics is, however, much wider. Copyright © 2004 Society of Chemical Industry     

Comparative proteome analysis of glutenin synthesis and accumulation in developing grains between superior and poor quality bread wheat cultivars

BACKGROUND: Wheat glutenins are the major determinants of wheat quality. In this study, grains at the development stage from three wheat cultivars (Jimai 20, Jin 411 and Zhoumai 16) with different bread‐making quality were harvested based on thermal times from 150 °C_d to 750 °C_d, and were used to investigate glutenin accumulation patterns and their relationships with wheat quality. RESULTS: High and low molecular weight glutenin subunits (HMW‐GSs and LMW‐GSs) were synthesised concurrently. No obvious correlations between HMW/LMW glutenin ratios and dough property were observed. Accumulation levels of HMW‐GSs and LMW‐GSs as well as 1Bx13 + 1By16 and 1Dx4 + 1Dy12 subunits were higher in superior gluten quality cultivar Jimain 20 than in poor quality cultivar Jing 411 and Zhoumai 16. According to the results of two‐dimensional gel electrophoresis, six types of accumulation patterns in LMW‐GSs were identified and classified. The possible relationships between individual LMW‐GSs and gluten quality were established. CONCLUSION: The high accumulation level of HMW‐GSs and LMW‐GSs as well as 1Bx13 + 1By16 and 1Dx4 + 1Dy12 subunits contributed to the superior gluten quality of Jimai 20. Two highly expressed and 16 specifically expressed LMW glutenin subunits in Jimain 20 had positive effects on dough quality, while 17 specifically expressed subunits in Zhoumai 16 and Jing 411 appeared to have negative effects on gluten quality. Copyright © 2011 Society of Chemical Industry     

The influence of 1B/1R chromosome translocation on gluten protein composition and technological properties of bread wheat

The Austrian bread wheat Amadeus without and with 1BL/1RS translocation and three further translocation genotypes with known HMW subunit compositions were grown under the same environmental conditions. Their flours were characterised by the determination of crude protein content and, partly, by the determination of glutathione and cysteine. Furthermore, the qualitative and quantitative composition of gluten protein types was analysed by a combined extraction and reversed phase HPLC procedure. Dough development time, maximum resistance and extensibility of dough and gluten, and bread volume were determined by means of microscale methods. Protein, glutathione and cysteine contents of flours were only slightly influenced by translocation. The HPLC patterns of gliadins and glutenin subunits showed that translocation caused characteristic changes concerning ω‐gliadins, γ‐gliadins and LMW subunits of glutenin. The amount of ω 1,2‐gliadins was significantly increased and that of LMW subunits decreased. The effect of translocation on the rheological properties of dough and gluten was characterised by a strongly reduced dough development time, reduced maximum resistance and increased extensibility. Bread volume was decreased by about 10%. The amount of glutenin subunits was correlated with dough development time, resistance of dough and gluten, and bread volume to a higher extent (r = 0.79–0.91) than the amount of gliadins (r = 0.52–0.80). Correlation coefficients for LMW subunits were higher (r = 0.82–0.88) than those for HMW subunits (r = 0.35–0.61) when all five wheats were included. Instead, when only translocation lines were considered, HMW subunits (r = 0.89–0.98) were more important than LMW subunits (r = 0.64–0.86). Altogether, the results demonstrate that translocation causes important quantitative as well as qualitative changes in gluten protein composition which can be efficiently determined by reversed phase HPLC. © 2000 Society of Chemical Industry     

Biotechnology of Wheat Quality

Wheat gluten proteins are largely responsible for the visco-elastic properties that allow doughs to be processed into bread and various other food products including cakes, biscuits (cookies), pasta and noodles. Detailed biochemical and biophysical studies are revealing details of the molecular structures and interactions of the individual gluten proteins, and their roles in determining the functional properties of gluten. In particular, one group of gluten proteins, the high molecular weight (HMW) subunits of glutenin, have been studied in detail because of their role in determining the strength (elasticity) of doughs. The development of robust transformation systems for bread wheat is now allowing the role of the HMW subunits to be explored experimentally, by manipulating their amount and composition in transgenic plants. Such studies should lead to improvement of the processing properties of wheat for traditional end uses and the development of novel end uses in food processing or as raw material for other industries. © 1997 SCI.     

The influence of nitrogen fertilisation on quantities and proportions of different protein types in wheat flour

The flours of 13 wheat varieties grown at different levels of nitrogen fertilisation were characterised by the quantitative determination of flour protein groups and gluten protein types using a combined extraction/HPLC procedure. The results demonstrate that the quantities of albumins and globulins were scarcely influenced by different nitrogen fertilisation, whereas those of gluten proteins (gliadins, glutenins) were strongly influenced. The effect on gliadins was more pronounced than on glutenins, as well as the effect on major protein types (α-gliadins, γ-gliadins, LMW subunits of glutenin) in comparison with minor types (ω-gliadins, HMW subunits of glutenin). The proportions of hydrophilic proteins (ω-gliadins, HMW subunits of glutenin) were increased by high levels of nitrogen and those of hydrophobic proteins (γ-gliadin, LMW subunits of glutenin) were decreased. The degree of the effects on both quantities and proportions of flour protein groups and gluten protein types was strongly dependent on the variety. © 1998 SCI.     

Wheat-bug damage in New Zealand wheats. Development of a simple SDS-sedimentation test for bug damage

Lines of three varieties of New Zealand wheat (cv. Aotea, Oroua and Kopara) damaged by wheat-bug were investigated. Hydrolysis of gluten proteins was shown to be of the endo- rather than the exo-proteolytic type. Electrophoresis revealed that the bug protease had a high specificity for the high molecular weight (HMW) glutenin subunits. An autolytic assay method was developed based on the decrease in sodium dodecyl sulphate (SDS)-sedimentation volume for bug-damaged flours when they were incubated in distilled water for 30 min at 37°C. This method was specific for bug damage and exhibited no interference from other grain defects such as heat damage, field sprouting and laboratory germination.     

不同类型专用小麦HMW-GS和GMP含量与面筋含量的关系

高分子质量谷蛋白质亚基(HMW-GS)是决定小麦加工品质的重要因子,以25个小麦品种为材料,采用SDS-PAGE电泳方法研究亚基组成、各位点(Glu-A1、Glu-B1、Glu-D1)亚基积累量、谷蛋白大聚合体(GMP)含量及其与面筋含量的关系.研究结果表明:低蛋白小麦品种HMW-GS组成主要以"N、7+8/7+9、2+12"亚基组合为主,高蛋白小麦品种HMW-GS组成则主要以"1、7+8/7+9、5+10"亚基组合为主,中等蛋白小麦品种兼有以上两种类型亚基组合;亚基组成相同的小麦,籽粒GMP含量和蛋白质含量仍有较大差异.相关分析表明,HMW-GS亚基总积累量与籽粒蛋白质含量和GMP含量呈极显著正相关,与干、湿面筋含量成正相关;GMP含量与干、湿面筋含量、蛋白质含量均呈极显著正相关,说明小麦品质类型虽与HMW-GS组成有关,但亚基积累强度不同可能是导致籽粒品质类型差异的重要因素之一.     

Importance of amounts and proportions of high molecular weight subunits of glutenin for wheat quality

 High molecular weight (HMW) subunits of wheat glutenin are generally considered to play a key role in gluten formation and structure, and to be closely related to wheat quality. Though quantities of HMW subunits in flour have been proposed to be as important for wheat quality as their structures, only few quantitative data are available in the literature. Therefore, two assortments of wheat consisting of 13 international and 16 German cultivars were analyzed for their contents and proportions of single HMW subunits using an extraction and HPLC procedure on a micro-scale. The results were compared with quantitative data from the literature that were obtained by sodium dodecylsulfate polyacrylamide gel electrophoresis combined with densitometry or by reversed-phase HPLC combined with UV detection. The quantitative analyses demonstrated that the contents of HMW subunits varied within a broad range dependent on genotype and growing conditions. The proportions of subunits within a given subunit combination, however, varied only within a small range. Generally, subunits 2, 5, 7, 10 and 12 were major components and subunits 1, 2*, 6, 8 and 9 were minor components. The levels of HMW subunits were highly correlated to dough development time, maximum resistance of dough and gluten, and bread volume. Among HMW subunits the x-type components (subunits 1–7) were much more important than the y-type components (subunits 8–12). In particular, the presence of subunit 5 (which has an additional cysteine residue) and of subunit 7 (which occurs in the greatest amounts) contributed to high wheat quality. Received: 22 June 1999     

Variation in concentrations of high-molecular-weight glutenin subunits and macropolymers in wheat grains of a recombinant inbred lines population and in two contrasting eco-sites in China

BACKGROUND: Concentrations of high‐molecular‐weight glutenin subunits and macropolymers in wheat grains are important indicators of grain quality, which are genetically determined and affected by environmental factors. The 6 VS·6AL translocation chromosome segment is reported to own high powdery mildew and yellow rust resistance genes of Pm21 and Yr26. This study investigated the variation in concentrations of high‐molecular‐weight glutenin subunits (HMW‐GS) and glutenin macropolymer (GMP) in response to the 6 VS·6AL translocation segment and the two contrasting sites. RESULTS: Large variations in concentrations of HMW‐GS and GMP were observed within lines containing different HMW‐GS compositions and between the contrasting eco‐sites. However, 6 VS·6AL chromosome translocation segment showed no significant effects on concentrations of HMW‐GS and GMP. In addition, HMW‐GS concentration was also found to be significantly correlated with the GMP concentration. CONCLUSION: Concentrations of HMW‐GS and GMP are largely affected by the eco‐sites and the composition of HMW‐GS, whilst not by the presence of 6 VS·6AL chromosome segment translocation. The 6 VS·6AL translocation is suggested as potential donor for breeding wheat cultivars for high resistence to powdery mildew and yellow rust with less risk of undesirable effects on grain quality. Copyright © 2012 Society of Chemical Industry     

Comparative investigations of gluten proteins from different wheat species I. Qualitative and quantitative composition of gluten protein types

In contrast to the hexaploid common (bread) wheat, little information is available on the qualitative and quantitative compositions of gluten proteins from other cultivated wheat species. Therefore, representatives of hexaploid spelt, tetraploid durum wheat and emmer, and diploid einkorn were compared with three classes of common wheat (winter wheat, spring wheat, wheat rye hybrid). The flours were extracted to yield total endosperm proteins and the gluten protein fractions (gliadins and glutenin subunits). The extracts were characterised using sodium dodecyl sulfate polyacrylamide gel electrophoresis and reversed-phase HPLC; both methods revealed that gluten protein groups and types known from common wheat (ω-, α-, γ-gliadins, HMW and LMW subunits of glutenin) were present in all species. The HPLC platterns of gliadins and glutenin subunits from species with the same genome composition (common wheat/spelt or durum wheat/emmer) were related, and those of einkorn quite different. According to the quantities determined by reversed-phase HPLC, α-gliadins were predominant in most cases, followed by γ-gliadins and LMW subunits; ω-gliadins and HMW subunits were generally minor components. Common wheats were characterised by the highest proportions of total glutenins and HMW subunits, which are known to be important for breadmaking quality. Moreover, the lower ratio of gliadins to glutenins was typical. Emmer had the lowest proportions of total glutenins and of HMW and LMW subunits, together with einkorn the highest proportion of α-gliadins, and, by far, the highest ratio of gliadins to glutenins. The values for spelt and durum wheat were mostly in a medium range between common wheats, emmer, and einkorn, respectively. Amongst common wheats, spring wheat was characterised by more balanced quantities of α- and γ-gliadins, and wheat rye hybrid by the highest proportions of ω-gliadins. Received: 26 November 1999     

高分子量谷蛋白亚基近等基因系面粉品质及其流变学特性的研究

分析了高分子量谷蛋白亚基近等基因系的高分子量谷蛋白亚基组成，并对该近等基因系进行了品质性状的分析，结果表明：Glu-D1位点的变化比Glu-B1位点的变化对面粉品质和流变学特性影响大；并且同时具有Glu-B1 14 15和Glu-D1 5 10的近等基因系2号无论是面粉品质还是流变学特性都优于其它两个近等基因系8号(null，7 9，5 10)和近等基因系13号(null，14 15，10)；而8号又优于13号，即Glu-B1 14 15的存在没有弥补Glu-D1 5亚基的缺失。     

Sulfur in wheat gluten: In situ analysis by X-ray absorption near edge structure (XANES) spectroscopy

The sulfur containing gluten proteins largely determine the baking quality of wheat. In order to probe the speciation of sulfur, gluten proteins [gliadin, high molecular weight (HMW) and low molecular weight (LMW) subunits of glutenin], stored glutenin subunits as well as flour were investigated in situ by S K-edge X-ray near edge absorption structure (XANES) spectroscopy. The spectra confirmed the existence of disulfide bonds in oxidised (oxygen stream) glutenin subunits, supporting their significance for the formation of gluten networks. Additionally, glutenin subunits, which were stored under ambient air and temperature conditions, predominantly contained sulfur of higher oxidation states (sulfoxide, sulfonic acid). The disulfide state and also sulfoxide and sulfonic acid states were detected after reoxidation of glutenin subunits with potassium bromate.     

Relationship between gluten degradation by Aelia spp and Eurygaster spp and protein structure

Gluten from wheat damaged by heteropterous insects loses its functionality after a short period of resting. In this study the properties of the gluten from damaged wheat are compared with that from sound wheat in order to understand the changes produced during incubation at 37 °C. The amounts of free thiol and amino groups were quantified, obtaining a marked increase of those groups during incubation of the damaged wheat. The thermal characterization of the damaged gluten showed a decrease in the denaturation temperature and a pronounced increase in the protein denaturation enthalpy after a short incubation, although the value of that enthalpy greatly dropped with a longer incubation period. The high‐molecular‐weight glutenin subunits (HMW‐GS) were rapidly hydrolysed while the low‐molecular‐weight glutenin subunits (LMW‐GS) showed a slower degradation. It seems that the HMW‐GS backbone was first hydrolysed, leading to a protein structure with higher thermal stability but, as the hydrolysis proceeded, a deeper degradation of the structure yielded a protein structure with lower denaturation enthalpy. The loss of gluten functionality results from complex changes in the gluten structure at the first and second level of the protein organization structure. Copyright © 2005 Society of Chemical Industry     

Dispersion in the Presence of Acetic Acid or Ammonia Confers Gliadin‐Like Characteristics to the Glutenin in Wheat Gluten

Spray‐dried gluten has unique properties and is commercially available in the food industry worldwide. In this study, we examined the viscoelastic properties of gluten powder prepared by dispersion in the presence of acetic acid or an ammonia solvent and then followed by lyophilization instead of a spray drying. Mixograph measurements showed that the acid‐ and ammonia‐treated gluten powders had marked decreases in the time to peak dough resistance when compared with the control gluten powder. The integrals of the dough resistance and bandwidth for 3 min after peak dough resistance decreased in both treated gluten powders. Similar phenomena were observed when gliadin was supplemented to gluten powders. Basic and acidic conditions were applied to the acid‐ and ammonia‐treated gluten powders, respectively, and the viscoelastic behaviors were found to depend on the pH in the gluten dispersion just before lyophilization. These behaviors suggest that gluten may assume a reversible change in viscoelasticity by a fluctuation in pH during gluten dispersion. SDS‐PAGE showed that the extractable proteins substantially increased in some polymeric glutenins including the low molecular weight‐glutenin subunit (LMW‐GS) when the ammonia‐treated gluten powder was extracted with 70% ethanol. In contrast, the extractable proteins markedly increased in many polymeric glutenins including the high molecular weight‐glutenin subunit and/or the LMW‐GS when the acid‐treated gluten powder was extracted with 70% ethanol. It thus follows that the extractability of polymeric glutenin to ethanol increases similarly to gliadin when gluten is exposed to an acidic or a basic pH condition; therefore, glutenin adopts gliadin‐like characteristics.     

Wheat gluten: high molecular weight glutenin subunits--structure, genetics, and relation to dough elasticity

ABSTRACT:  Gluten proteins, representing the major protein fraction of the starchy endosperm, are predominantly responsible for the unique position of wheat amongst cereals. These form a continuous proteinaceous matrix in the cells of the mature dry grain and form a continuous viscoelastic network during the mixing process of dough development. These viscoelastic properties underline the utilization of wheat to prepare bread and other wheat flour based foodstuffs. One group of gluten proteins is glutenin, which consists of high molecular weight (HMW) and low molecular weight (LMW) subunits. The HMW glutenin subunits (HMW-GS) are particularly important for determining dough elasticity. The common wheat possesses 3 to 5 HMW subunits encoded at the Glu-1 loci on the long arms of group 1 chromosomes (1A, 1B, and 1D). The presence of certain HMW subunits is positively correlated with good bread-making quality. Glutamine-rich repetitive sequences that comprise the central part of the HMW subunits are actually responsible for the elastic properties due to extensive arrays of interchain hydrogen bonds. Genetic engineering can be used to manipulate the amount and composition of the HMW subunits, leading to either increased dough strength or more drastic changes in gluten structure and properties.     

Disulphide bonds in wheat gluten: further cystine peptides from high molecular weight (HMW) and low molecular weight (LMW) subunits of glutenin and from γ-gliadins

Glutenin was prepared from gluten of the wheat variety Rektor by extraction of gliadin with aqueous ethanol. It was cleaved successively into soluble peptides by the enzymes trypsin and thermolysin. Separation of the peptide mixtures was performed by gel permeation chromatography (GPC) on Sephadex G25 and reversed phase high performance liquid chromatography (RP-HPLC) on ODS-Hypersil. Cystine peptides were detected by differential chromatography of the samples prior to and after reduction. After isolation by multi-step RP-HPLC, the cystine peptides were reduced. The resulting cysteine peptides were alkylated with 4-vinylpyridine, separated by RP-HPLC and sequenced by means of the Edman degradation. The isolated cystine peptides represented a considerable portion of the total cysteine in glutenin: four out of seven cysteine residues of HMW subunits, and eight out of nine cysteine residues of LMW subunits are documented by at least one cystine peptide. Most of the peptides corresponded to known sequences of gluten protein components. From the structures of some tryptic peptides, inter- and intramolecular disulphide bonds for HMW subunits of glutenin have been proven. Cystine peptides from the thermolytic digest have been assigned to LMW subunits of glutenin and toγ-gliadins. Other peptides have been closely related to partial sequences of these protein components. The results have allowed several conclusions about the arrangement of intra- and intermolecular disulphide bridges in gluten proteins.