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J A Bietz

Publications and source records attributed to J A Bietz.

9 recordsLinked to original sources
PubMed2000

Acetonitrile as a buffer additive for free zone capillary electrophoresis separation and characterization of maize (Zeamays L. ) and sorghum (Sorghum bicolor L. Moench) storage proteins.

An improved method for separating and characterizing maize (Zea mays L.) and sorghum (Sorghum bicolor L. Moench) storage proteins by free zone capillary electrophoresis (FZCE) was developed. Previous electrophoretic methods for analyzing these proteins required high concentrations of urea to maintain protein solubility during separation. To overcome disadvantages of urea, we developed a FZCE method that mimicked reversed-phase high-performance liquid chromatography (RP-HPLC) in that it used high levels of acetonitrile (ACN) at low pH. The optimized FZCE buffer system consisted of 80 mM phosphate-glycine buffer, nominal pH 2.5, containing 60% ACN and a cellulose derivative to dynamically coat capillary walls. Resolution was similar to or higher than that previously achieved by FZCE buffers utilizing 8 M urea as a buffer additive. ACN concentrations of at least 50% were necessary to achieve acceptable separations; this ACN concentration is approximately that necessary to extract these storage proteins. ACN was equally effective as traditional ethanol solvents and 8 M urea for solubilizing maize and sorghum proteins. The ACN-based FZCE buffer system gave high repeatability (<0.3% relative standard deviation, measured over 15 consecutive injections) for migration time. Subclasses of maize and sorghum storage proteins were identified, and genotypes of each cereal were successfully differentiated using ACN-containing buffers. This FZCE method may be applicable for the analysis of other hydrophobic proteins without the use of urea.

Acetonitriles↗
PubMed1998

High-performance capillary electrophoresis of cereal proteins.

Cereal grains are widely used of human foods and animal feed throughout the world. Cereals provide dietary protein, which also often has a functional role, as wheat gluten does in bread. Cereal proteins are unique in many ways: they are highly complex and heterogeneous, are often difficult to extract, and aggregate readily, making them difficult to characterize. Because of the economic importance and widespread use of cereal proteins, however, many techniques have been used for their analysis. High-performance capillary electrophoresis (HPCE) is one of the newest techniques to be so used. This review describes the development of charge- and size-based HPCE methods for analysis of cereal grain proteins, and the use of these methods for cultivar identification, classification, and prediction of quality. HPCE is versatile, rapid, easily automated, readily quantified, and provides high-resolution separations. Clearly, HPCE is a valuable addition to other methods of cereal protein analysis and should, in time, be applicable to all protein classes from all cereals.

Edible Grain↗
PubMed1992

Electrophoresis and chromatography of wheat proteins: available methods, and procedures for statistical evaluation of the data.

Analysis of gluten proteins from the wheat grain endosperm has long challenged the analytical chemist. Several hundred unique polypeptides are present, many in large polymers. This complexity, plus useful relationships of composition to genotype and quality, encouraged development and application of electrophoresis and chromatography for gluten analysis. We review the methods of polyacrylamide gel electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing and high-performance liquid chromatography available for study of wheat proteins. Singly and in combination, they provide rapid, reproducible, high-resolution separations based on size, charge, or surface hydrophobicity. As challenging and important as the analyses themselves, however, is interpretation of data. Subjective evaluation is sometimes possible, but statistical methods such as similarity scores, clustering, principal components, multiple linear regression, and partial least squares now are increasingly used for data analysis. We review the use of these procedures, and precautions necessary to avoid misinterpretation of data. Optimal evaluation of protein analytical data will enhance the value of such analyses in wheat breeding, marketing, and processing.

Chromatography↗
PubMed1983

Separation of cereal proteins by reversed-phase high-performance liquid chromatography.

Cereal proteins have been extremely difficult to purify and characterize owing to their heterogeneity, poor solubility and tendency to polymerize. High-performance liquid chromatography (HPLC) on a 300 A reversed-phase (RP)(C18) support (Syn Chropak RP-P), using acetonitrile as organic modifier in the presence of trifluoroacetic acid, has been found to be capable of high-resolution separations of these proteins; the resolution is often better than that obtained by any other chromatographic or electrophoretic method. Examples are presented showing separations of low-molecular-weight gliadins, omega-gliadins and ethanol-soluble reduced glutenin subunits from wheat and of zein from corn. In addition, proteins may be directly extracted from ground single kernels and subsequently analyzed by RP-HPLC; applications in genetic studies, in breeding programs and in varietal identification are proposed. In addition to its high resolution, RP-HPLC is superior to most other methods in speed, sensitivity, reproducibility and suitability for quantitation. Polypeptide chains of molecular weight up to 133,000 are recovered in high yields and the column capacity is high, demonstrating that RP-HPLC is suitable for both preparative and analytical separations of proteins. RP-HPLC resolves proteins primarily on the basis of differences in surface hydrophobicity, so it therefore complements, rather than duplicates, other techniques that separate proteins on the basis of size or charge. RP-HPLC promises to become an invaluable technique for the fractionation and characterization of proteins from cereals and other sources.

Chromatography, High Pressure Liquid↗
PubMed1982

Cereal prolamin evolution and homology revealed by sequence analysis.

Prolamin mixtures were isolated from oats, rice, normal and high-lysine sorghum, two varieties of pearl millet, two strains of teosinte, and gamma grass and subjected to NH2-terminal amino acid sequence determinations. In each case (except for rice, whose prolamins apparently have blocked or unavailable NH2-terminal residues), primarily a single sequence was observed despite significant heterogeneity, suggesting that prolamin homology in each cereal arose through duplication and mutation of a single ancestral gene. Comparisons were then made to prolamin sequences previously determined for wheat, corn, barley, and rye. Within genera, different varieties or subspecies exhibited few differences, but more distantly related genera, subtribes, and tribes showed increasingly large differences. Within the subfamily Festucoideae, no homology was apparent between prolamins of oats and those of the subtribe Triticinae (including wheat, rye, and barley, for which prolamin homology was previously demonstrated). Within the subfamily Panicoideae, corn was shown to be closely related to teosinte but more distantly to Tripsacum. Sorghum was shown to have diverged less from corn than had millet. These comparisons demonstrate that prolamin sequence analyses can successfully predict and clarify evolutionary relationships of cereals.

Amino Acid Sequence↗
PubMed1977

Disulfide bonds: key to wheat protein functionality.

Disulfide bonds in wheat proteins are major factors that determine the properties of the proteins and their functionality in wheat flour. The gliadin proteins contain mostly intramolecular disulfide bonds. In contrast, the high-molecular-weight glutenins are formed by disulfide linkages of several diverse polypeptide chains which have been separated and characterized. The linkage of these proteins in a fairly linear array contributes to the unique viscoelastic properties of glutenin. The glutenin has been separated into two fractions differing in molecular weight. The amount of highest molecular weight component is correlated with the rheological behavior of the flours from different wheat varieties. Various oxidizing and reducing agents are widely used to alter the functional behavior of wheat proteins by the action on sulfhydryl and disulfide groups.

Amino Acids↗