Analysis and properties of arabinoxylans from discrete corn wet-milling fiber fractions.

Three fibrous corn wet-milling fractions, coarse fiber, fine fiber, and spent flake, were isolated. More highly valued uses are sought for these milling products, which are generally directed into the corn gluten feed product stream. Coarse fiber was further dissected into pericarp and aleurone layers. An alkaline hydrogen peroxide process was used to efficiently extract corn fiber gum (CFG) from each of the materials. CFG is a hemicellulose B arabinoxylan which also contains low levels of D,L-galactose and D-glucuronic acid. CFG yield information was obtained from each source, as well as structural information in terms of degrees of branching of the beta-D-xylopyranose backbone with alpha-L-arabinofuranosyl moieties. There were significant differences in degree of branching among the CFGs from the various fractions. A novel capillary electrophoresis procedure was developed to measure these differences. Solution viscosity differences among the CFGs were also observed.

Purification of commercial gellan to monovalent cation salts results in acute modification of solution and gel-forming properties.

Lithium, sodium, potassium, and ammonium salts of the industrial polysaccharide gellan were prepared. The salts were freely soluble in water at room temperature (25 degrees C). The opinion had been generally held that heating to 100 degrees C was necessary for gellan to achieve complete solubility in the presence of mono- or multivalent cations. Then, upon cooling, the solutions would form gels. These conclusions were based on the properties imposed upon gellan samples by the presence of contaminating divalent cations. Commercial gellan samples contain calcium and magnesium at levels exceeding 0.9%, sufficient for counterion formation with over one-third of gellan's carboxyl groups. Purification was rapid and included sequential treatments with a cation-exchange (H+) resin, LiOH, NaOH, KOH, or NH4OH, and an anion-exchange (Cl-) resin. About 95% of the divalent cations and nearly 90% of the phosphate that contaminated commercial gellan were removed. The purified monovalent salts of gellan set in the presence of divalent cations and provide well-defined agents for gelling media used for propagation of microbes and plants. In a manner analogous to sodium alginate, solutions of lithium, sodium, potassium, or ammonium gellanate form beads when dropped into solutions of divalent cations. This property was exploited for entrapment of enzymes and cells in beads.

Partitioning of Intermediary Carbon Metabolism in Vesicular-Arbuscular Mycorrhizal Leek.

Vesicular-arbuscular mycorrhizal fungi are symbionts for a large variety of crop plants; however, the form in which they take up carbon from the host is not established. To trace the course of carbon metabolism, we have used nuclear magnetic resonance spectroscopy with [13C]glucose labeling in vivo and in extracts to examine leek (Allium porrum) roots colonized by Glomus etunicatum (and uncolonized controls) as well as germinating spores. These studies implicate glucose as a likely substrate for vesicular-arbuscular mycorrhizal fungi in the symbiotic state. Root feeding of 0.6 mM 1-[13C]glucose labeled only the fungal metabolites trehalose and glycogen. The time course of this labeling was dependent on the status of the host. Incubation with 50 mM 1-[13C]glucose caused labeling of sucrose (in addition to fungal metabolites) with twice as much labeling in uncolonized plants. There was no detectable scrambling of the label from C1 glucose to the C6 position of glucose moieties in trehalose or glycogen. Labeling of mannitol C1,6 in the colonized root tissue was much less than in axenically germinating spores. Thus, carbohydrate metabolism of host and fungus are significantly altered in the symbiotic state.

Binding geometry, stoichiometry, and thermodynamics of cyclomalto-oligosaccharide (cyclodextrin) inclusion complex formation with chlorogenic acid, the major substrate of apple polyphenol oxidase.

The inclusion complexes of cyclomaltohexaose (alpha-CD), cyclomaltoheptaose (beta-CD), cyclomaltooctaose (gamma-CD), and polymerized beta-CD (beta-CDn) with chlorogenic acid (CA), the major substrate of apple fruit polyphenol oxidase (PPO), were studied with regard to pH, ionic strength, and temperature in model buffer systems and apple juice. The thermodynamics of CD.CA inclusion complex formation, which were studied in solution using UV spectrophotometry, displayed enthalpy-entropy compensation typical of processes driven by solvation phenomena. We also found that the apparent association constants (K) of the CD.CA equilibrium were relatively insensitive to pH for beta-CD, compared to alpha- and gamma-CDs, but were subject to substantial enhancement at low ionic strengths. The beta-CD.CA inclusion complex was also characterized for binding geometry and stoichiometry at 9.4 T and 25 degrees C in 0.05 M Na phosphate buffer by 1H NMR spectroscopy. A 1:1 stoichiometric ratio for the complex was found using the method of continuous variations. 1H Spin-lattice relaxation and chemical-shift data indicate that the phenolic ring of CA docks within the cavity of beta-CD. The Ks for beta-, alpha-, and gamma-CD determined in apple juice, which contains a mixture of PPO substrates, were found to correlate with PPO activity-related data. Apple juice, treated with beta-CDn, did not brown until CA was added back. These latter findings strongly argue that the mechanism for inhibition of juice browning with cyclodextrins was mainly due to the binding of PPO substrates and not some other means such as enzyme inactivation via sequestration of Cu2+ by CDs.

Assay of reducing end-groups in oligosaccharide homologues with 2,2'-bicinchoninate.

For the first time, reducing values of homologous series of oligosaccharides have been determined by the 2,2'-bicinchoninate assay. The extent of Cu2+ reduction was monitored by spectrophotometric measurement of Cu+/2,2'-bicinchoninate complexes. Conditions of the assay were optimized so that relatively uniform reducing values were obtained with oligosaccharides derived from starch, polygalacturonic acid, and chitin, regardless of degree of polymerization. The uniform values resulted because members of each oligosaccharide series were oxidized to similar levels beyond the aldonic acid stage, while oxidation was limited to the reducing-terminal residue. This conclusion was based on direct measurements of quantitative loss of paramagnetic copper (Cu2+) by electron paramagnetic resonance spectroscopy.

Preparative liquid chromatography of carbohydrates: mono- and di-saccharides, uronic acids, and related derivatives.

The general principles and practical aspects of preparative high-performance liquid chromatography (l.c.) of mono- and di-saccharides, sugars acids, lactones, and N-acetylated amino sugar derivatives are described. Milligram to gram quantities of these carbohydrates were isolated on semi-preparative (0.78 X 30 cm) or preparative (approximately 2.0 X 30 cm) columns packed with aminopropyl silica gel provided better resolution of individual mono- and di-saccharides, but columns of cation-exchange resin had higher capacity and were more durable and economical to use. Preparative, cation-exchange columns were operated at flow rates of less than 5 mL/min and pressures of approximately 1-2 MPa, allowing them to be used on unmodified analytical l.c. systems. Details are given for the efficient packing, use, and care of these columns, and on the effects of column selectivity, packing technique, and sample size on chromatographic resolution. Isolation of naturally occurring sugars from biological sources on a laboratory-packed column is described.

Mass spectrometric 13C/12C determinations to detect high fructose corn syrup in orange juice: collaborative study.

The 13C/12C ratios in orange juice are sufficiently uniform and different from those in high fructose corn syrup (HFCS) so that the addition of HFCS to orange juice can be detected. HFCS averages -9.7% (parts per thousand) delta 13C, orange juice averages -24.5%, and mixtures of HFCS and orange juice possess intermediate values. One pure orange juice and 4 orange juice -HFCS mixtures containing from 25 to 70% orange juice were properly classified by 7 collaborators. Samples with delta 13C values less negative than -22.1%, 4 standard deviations from the mean of pure juices, can, with a high degree of confidence, be classified as adulterated. Samples with values more negative than -22.1% must be considered unadulterated with HFCS, because pure orange juices possess a range of delta 13C values. The 13C/12C mass spectrometric method was adopted official first action for detecting HFCS in orange juice.

Charcoal column/thin layer chromatographic method for high fructose corn sirup and spectrophotometric method for hydroxymethylfurfural in honey: collaborative studies.

A new spectrophotometric method is described for determining hydroxymethylfurfural in honey in which interfering background absorption of honey is corrected for by use of a bisulfite-treated sample as blank. Two procedures for detecting high-fructose corn sirup (HFCS) in honey were also tested. In one, charcoal column pretreatment is used to concentrate trace oligosaccharides, followed by thin layer chromatography to differentiate those of HFCS from those of honey. The other method depends on measurement of the isomaltose/maltose ratio by gas-liquid chromatography. The charcoal/thin layer chromatographic method for HFCS has been adopted official first action. The bisulfite method for hydroxymethylfurfural has been adopted interim first action.

Gas-liquid chromatographic test for honey adulteration by high fructose corn sirup.

A gas-liquid chromatographic (GLC) method has been developed to detect the addition to honey of high fructose corn sirup (HFCS). Samples are derivatized directly with trimethylsilane, cholestane is added as an internal standard, and the levels of maltose (includes other minor disaccharides) and isomaltose are determined after chromatography on OV-17. Domestic and imported honey samples (115) contained 2.00% maltose and 0.71% isomaltose. HFCS samples (21) contained 1.50% maltose, and 2.09% isomaltose. A discriminatory equation was developed (D = 2.73 - 5. 35 (Isomaltose/maltose)) and, when applied to the data for these samples and 37 adulterated samples, 81.4% of authentic honey samples and 78.4% of samples known to be adulterated with HFCS were correctly classified.

Carbon-13/carbon-12 ratio is relatively uniform among honeys.

The variability of the carbon-13/carbon-12 ratio in honeys was evaluated preliminary to use of the ratio to detect the addition of high-fructose corn syrup to honey. Eighty-four honey samples representing 34 states and including 37 floral types from 17 plant families were analyzed. The mean value of the per mil increment in carbon-13 (delta13C) for all samples is -25.2 per mil, and the coefficient of variation is 3.7 percent. This is the smallest variation yet encountered for a honey constituent or physical property. The range and magnitude of the values suggest that the floral sources are C(3) plants.