Fractionation of secalins and hordeins by preparative electrophoresis at acid pH

Rumbo, Martı́n; Margheritis, Analia Ines; Chirdo, Fernando Gabriel; Giorgieri, Sergio A.; Fossati, Carlos A.; Añón, Marı́a Cristina

doi:10.1007/s00217-001-0441-6

Cited by 2 publications

(2 citation statements)

References 8 publications

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“…In report Rumbo et al (2002), the optimization of a preparative electrophoretic method to fractionate secalins is described. Separation was performed in preparative 7% polyacrylamide gels of 4 cm length at pH 3.1.…”

Section: Resultsmentioning

confidence: 99%

Separation Of Rye Secalins By A-Page

Petrovičová¹,

Balážová²,

Vivodík³

et al. 2017

Agrobiodiversity for Improving Nutrition, Health and Life Quality

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The aim of our study was to evaluate the electrophoretic profiles of secalins of fifteen genotypes of rye (Secale cereale L.), which were obtained by polyacrylamide gel electrophoresis in the presence at pH 3.1 (A-PAGE). Electrophoretic separation of storage proteins was conducted according to the methodology recommended by an international organization ISTA, with some own modifications. Fractions from the preparative separation were pooled in such a way that no components from one pool were present in the others. Doc-It LS software was used to detect and to calculate variability of genotypes within individual rye species.Preparative electrophoresis at low pH allowed a simple separation of γ75k-secalins, ω-secalins and γ40k-secalins from the crude material under non-denaturing conditions. The content of γ75k-secalins varied in analyzed collection of rye from 22.86% (variety Čerkascanka tetra) to 53.93% (genotype Valtické) with an average 39.27%. The proportion of ω-secalins in our samples was in an average 39.27%. The largest percent representation of ω-secalins was proved in variety České (58.68%) and the lowest part of this subunits was detected in variety Radomske (25.83%). Our results showed that average representation of γ40k-secalins was 22.05%, with the highest content in variety Vígľašské (32.44%) overleaf with the lowest part of this fraction was detected in Valtické (8.28%). The variety of wheat Chinese spring and Marquis were used as standards. Storage proteins consist of three fractions, which are the main part of grain proteins and are used as a marker not only for genetic variability investigation, but also for characterization of genotypes. These pooled fractions could be used as starting material for single polypeptide purification. Keywords: cereals; Secale cereale; electrophoresis; A-PAGE; secalins IntroductionRye (Secale cereale L) is a traditional cereal in Central, Eastern and Northern Europe, where it is used for the production of bread and crispbread and as fodder (Hansen et al., 2003). The cereal proteins contribute to the nutritional value of the diet and they are integral and fundamental part of food components. Nutritionally, they are the good source of energy and its amino acids are essential for growth and maintenance (Horszwald et al., 2009). Functionally, they affect the physicochemical and sensory properties of various foods. Recently published data indicate that rye and rye-based products (including breads) are a good source of lignans, phytosterols and phenolic compounds that are biologically active and posses antioxidant properties (mainly they are good free-radical scavengers, reducing agents, potential complexers of prooxidant metals and quenchers of the singlet oxygen formation) (Zielinski, 2002;Michalska et al., 2007;Horszwald et al., 2009 379Genetic diversity has played a vital role in the success of crop improvement. Knowledge of genetic diversity has been successfully used for efficient germplasm management and utilization, genetic fingerprinting and genotype selection ...

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Section: Resultsmentioning

confidence: 99%

Separation Of Rye Secalins By A-Page

Petrovičová¹,

Balážová²,

Vivodík³

et al. 2017

Agrobiodiversity for Improving Nutrition, Health and Life Quality

View full text Add to dashboard Cite

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“…Protein content is a key characteristic used for accepting or rejecting barley for malting, and therefore it is important to have methods that can separate, characterize, and identify barley proteins. The two most commonly used identification techniques, polyacrylamide gel electrophoresis (PAGE) and high-performance liquid chromatography (HPLC), have recently been reviewed ( − ). Matrix-assisted laser desorption ionization (MALDI)−time of flight mass spectrometry (TOF-MS) also seems to be a useful alternative technique for the identification of these barley prolamins, with a detection sensitivity of about 50−100 ng total protein loaded ().…”

Section: Introductionmentioning

confidence: 99%

Free-Zone Capillary Electrophoresis Analysis of Hordein Patterns at Different Stages of Barley Malting

Garcı́a-Villalba

Cortacero-Ramírez

Segura‐Carretero

et al. 2006

J. Agric. Food Chem.

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We have carried out a comparison of hordein patterns at different stages of the malting process using free-zone capillary electrophoresis (FZCE). FZCE has proved to be a suitable technique for the separation and characterization of hordeins in barley seeds. Assays of protein extraction and electrophoretic procedures led us to conclude that hordeins were best extracted with 40% ethanol and analyzed using 50 mM phosphate-glycine, pH 2.5, containing 20% ACN and 0.05% HPMC, at 12.5 kV and 45 degrees C, with 10 s hydrodynamic injection at 0.5 psi and 50 microm i.d. x 31 cm uncoated fused-silica capillary. Our results afford useful information about changes in the composition of these proteins in barley during malting.

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Fractionation of secalins and hordeins by preparative electrophoresis at acid pH

Cited by 2 publications

References 8 publications

Separation Of Rye Secalins By A-Page

Separation Of Rye Secalins By A-Page

Free-Zone Capillary Electrophoresis Analysis of Hordein Patterns at Different Stages of Barley Malting

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