A simple extraction and one-step SDS–PAGE system for separating HMW And LMW glutenin subunits of wheat and high molecular weight proteins of rye

Zhen, Zhu; Mares, Daryl J.

doi:10.1016/s0733-5210(09)80057-x

Cited by 17 publications

(10 citation statements)

References 16 publications

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“…Wieser et al (1994) and Fu and Sapirstein (1996a) demonstrated that 50% 1-propanol could effectively extract all monomeric proteins (albumins, globulins, and gliadins) from flour. Several other articles have also reported this, as well as the fact that aqueous alcohols can also extract soluble polymeric proteins (Kruger et al 1988, Marchylo et al 1989, Singh et al 1991, Zhen and Mares 1992, Fu and Sapirstein 1996a. Based on these earlier works, we tested three different alcohols at a wide range of concentrations, as well as acetic acid and SDS, for their ability to remove polymeric and monomeric proteins.…”

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confidence: 74%

A Rapid Method for Quantitation of Insoluble Polymeric Proteins in Flour

et al. 1998

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Cereal Chem. 75(3):374-379The baking properties of several genotypes of U.S. hard wheats grown in state nurseries for the Wheat Quality Council (WQC) were analyzed by the Hard Winter Wheat Quality Laboratory. Flours (250 mg) from each individual line and location were extracted three times with 50% 1propanol (1 mL) for 5 min each. Samples were vortexed continually during extraction. This method was effective in removing most monomeric proteins. Negligible detectable protein was found in the third extract. Significant amounts of polymeric glutenin were also extracted. Pellets were oven-dried (130°C) for 1 hr and analyzed for protein content using nitrogen combustion analysis. Protein remaining in the pellet consisted mainly of polymeric protein. The amount of gliadin and soluble polymeric protein could also be measured by separating the supernatant by size-exclusion chromatography. Good correlations between dough strength parameters and amounts of pellet protein and the relative amount of pellet protein (pellet protein/flour protein) were found for all samples. This procedure was simple and rapid, with the potential of analyzing large numbers of samples per day with good reproducibility.

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confidence: 74%

A Rapid Method for Quantitation of Insoluble Polymeric Proteins in Flour

et al. 1998

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“…Proteins from 300-mg samples of wheat flour and extrudates (ground to 40 mesh) were extracted with a 12 mL of solution of 70% aqueous ethanol, 1% SDS, and 5% β-mercaptoethanol by magnetic stirring for 24 hr at room temperature. This procedure was adapted from Zhen and Mares (1992). The mixture of solvents commonly used with wheat proteins was found in preliminary studies to be more effective in extracting total protein from extrudates than SDS/β-mercaptoethanol alone, and coextraction of starch was minimized.…”

Section: Methodsmentioning

confidence: 99%

“…Wheat flours were extracted sequentially with 0.1N NaCl and 70% ethanol to isolate albumins and globulins, and then gliadins, respectively. The remaining material was treated with 70% ethanol, 1% SDS, and 5% 2-mercaptoethanol to extract glutenins (Zhen and Mares 1992). Proteins in extracts were isolated by precipitation in cold acetone.…”

Section: Methodsmentioning

confidence: 99%

Extrusion Chemistry of Wheat Flour Proteins: II. Sulfhydryl‐Disulfide Content and Protein Structural Changes

Rebello

Schaich

1999

Cereal Chem

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Cereal Chem. 76(5):756-763Effects of twin-screw extrusion conditions on wheat flour proteins were studied, using a two-level fractional factorial experimental design (11 and 14% protein content, 160 and 185°C, 16 and 20% moisture, 300 and 500 rpm screw speed, mass flow rate of 225 and 400 g/min). Total protein detectable by solid-phase bicinchoninic acid assay decreased slightly after extrusion, with greatest protein loss at 16% moisture and 160°C. Sulfhydryl content of both flours increased after extrusion at 185°C and 16% moisture with moderate specific mechanical energy (SME ≈ 400-600 kJ/kg) or 160°C and 16% moisture with high SME (SME > 1,000 kJ/kg). Disulfide bonds increased under comparable conditions but with moderate shear (SME = 510-540 kJ/kg). At 20% moisture and either temperature, sulfhydryl and total thiol contents decreased without corresponding increases in disulfides. Reversed-phase HPLC indicated gliadins were the fractions most affected by extrusion; high molecular weight glutenin subunits also were affected. Changes in gliadins were extensive at 185°C and 16% moisture and were minimal at 160°C and 20% moisture. SDS-PAGE confirmed the disappearance of protein bands and appearance of new material at low and high molecular weights, presumably resulting from polypeptide fragmentation followed by random radical recombination. Both protein fragmentation and cross-linking appeared to involve free radicals.Extrusion technology plays a central role in modern cereal-based industry, and significant advances in engineering have led to production of a wide range of extruded products. Nevertheless, surprisingly little still is known about the basic molecular phenomena that contribute to texturization and other product characteristics during extrusion of wheat flour products.Early studies on extrusion of wheat flour suggest that the mechanical and textural properties of extrudates are largely determined by gluten proteins in dough (Hauck 1980). The mechanical strength of extrudates is directly related to the gluten content of flour (Linko et al 1981). Faubion and Hoseney (1982a,b) extruded wheat starch with varying amounts (1-16%) of added wheat gluten or soy protein isolate and showed that the presence of protein is critically important for texturization and that the amount and type of protein in wheat flour markedly affects the textural properties of extrudates. Expansion decreased steadily as gluten protein content increased from 1 to 11%; at levels higher than 11%, expansion again increased. Shear and break strengths changed correspondingly. Similar effects were produced when yeast protein concentrate was extruded with wheat starch (Lai et al 1985). However, addition of soy proteins had the opposite effect: increasing expansion with protein levels of up to 8% and decreasing expansion with protein levels higher than 10%. Extrudates containing soy protein showed substantially greater expansion than those containing wheat gluten.These results indicate that both the type and level of protein exert important e...

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“…Both parents are soft grain types. The Glu-B1x subunit of CY12 had a slightly greater mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) than Glu-B1x from Chinese Spring (data not shown) and is numbered 7* according to earlier reports by Ng et al (1989), Pogna et al (1989) and Zhen and Mares (1992). Similarly, the Glu-B1 allele of CY12 is denoted as Glu-B1u.…”

Section: Plant Materialsmentioning

confidence: 94%

Effect of high-molecular-weight glutenin allele, Glu-B1d, from synthetic hexaploid wheat on wheat quality parameters and dry, white Chinese noodle-making quality

Tang¹,

Yang²,

Wu³

et al. 2010

Crop Pasture Sci.

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Abstract. Synthetic hexaploid wheat (SHW) represents a valuable source of new resistances to a range of biotic and abiotic stresses. Exploitation of these resistances in bread wheat breeding programs, however, is not necessarily straightforward and requires an assessment of potential negative effects on quality particularly from the genomes contributed by the durum parents used in the development of SHW. In particular, high-molecular-weight glutenin subunits (HMW-GS) 6+8 that are common in durum and SHW but, in bread wheat, are present at only a very low frequency in Chinese wheat cultivars and landraces and as a result there is only limited data on the effects of HMW-GS 6+8 on wheat processing quality and especially on dry, white Chinese noodles (DWCN). In this study, 131 recombinant inbred lines (RIL) were developed from a cross between a CIMMYT SHW 'Syn-CD780' and an elite Sichuan common wheat cultivar 'ChuanYu12'.The aim of this study was to investigate the effect of the HMW glutenin allele, Glu-B1d (6+8), from SHW on quality-related characteristics and DWCN making quality compared with the alternate allele Glu-B1u (7*+8). The RIL and parents were grown in three environments and analysed for 21 quality and noodle test parameters. Results showed the effect of Glu-B1d depended on both the parameters tested and glutenin subunit background contributed by alleles at the Glu-A1 and Glu-D1 loci. RIL with the Glu-B1d allele v. those with the Glu-B1u had significantly higher Zeleny sedimentation volume and falling number in the subunit backgrounds Glu-A1c/Glu-D1a and Glu-A1c/Glu-D1ah, significantly higher L* of dry flour in the background Glu-A1a/Glu-D1a; significantly higher dough development time, dough stability time, breakdown time and lower softness in both backgrounds Glu-A1c/Glu-D1a and Glu-A1c/GluD1ah; significantly higher values of most rapid visco analysis parameters, especially pronounced in the background GluA1c/Glu-D1a. The RIL with the Glu-B1d allele also showed significantly higher (P < 0.05) noodle total score (NTS) in the Glu-A1a/Glu-D1a background and significantly higher (P < 0.01) NTS and most components of sensory assessment in the Glu-A1c/Glu-D1a background. Overall, the results indicate that the allele Glu-B1d, 6+8, from synthetic hexaploids could, in general, have a positive influence on most bread wheat quality parameters and DWCN noodle-making, particularly when combined with particular glutenin subunits at Glu-A1 and Glu-D1.Additional keywords: high-molecular-weight glutenin subunit, recombinant inbred line, synthetic hexaploid wheat.

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A simple extraction and one-step SDS–PAGE system for separating HMW And LMW glutenin subunits of wheat and high molecular weight proteins of rye

Cited by 17 publications

References 16 publications

A Rapid Method for Quantitation of Insoluble Polymeric Proteins in Flour

A Rapid Method for Quantitation of Insoluble Polymeric Proteins in Flour

Extrusion Chemistry of Wheat Flour Proteins: II. Sulfhydryl‐Disulfide Content and Protein Structural Changes

Effect of high-molecular-weight glutenin allele, Glu-B1d, from synthetic hexaploid wheat on wheat quality parameters and dry, white Chinese noodle-making quality

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