Impact of annealing on the hierarchical structure and physicochemical properties of waxy starches of different botanical origins.

The effect of annealing on structure and physicochemical properties of four different waxy starches were investigated to understand the mechanism of annealing in the absence of amylose. Granule morphology, X-ray pattern, molecular order, and gelatinization enthalpy remained unchanged upon annealing. Relative crystallinity of waxy corn starch increased from 42.4 to 46.1% on annealing, while it remained unchanged in other starches. Annealing increased the gelatinization temperatures, while it decreased the temperature range. Waxy potato starch showed the highest increase in To, where it was 8.1 °C after 72 h of annealing. Viscosity profiles were only slightly affected by the annealing treatment. Susceptibility towards amylolysis increased in waxy corn and waxy barley, while it decreased in waxy potato and waxy rice. This study demonstrated that even in the absence of amylose, annealing caused changes in the structure and properties, however, the extents of these changes varied depending on the botanical origin.

Probing the microheterogeneity of O-specific chains from Yersinia ruckeri using capillary zone electrophoresis/electrospray mass spectrometry.

The analysis of underivatized oligosaccharides arising from mild acid hydrolysis of endotoxins from Yersinia ruckeri serotype O2 was achieved using on-line capillary zone electrophoresis-electrospray mass spectrometry (CZE-ESMS). This technique provided unparalleled resolution of the different glycans obtained from purified fractions of the native endotoxins or from hydrolysis of lipopolysaccharides from Y. ruckeri. Electrophoretic conditions enabling the separation of anionic and cationic analytes were developed to determine possible sites of heterogeneity on either the core or the O-chain glycans. Structural characterization of underivatized oligosaccharides identified in the ion electropherograms was achieved using tandem mass spectrometry under low-collision energy conditions.

Synthesis of glycoconjugates derived from various lipopolysaccharides of the Vibrionaceae family.

Conjugation of simple ketoses (such as 3-deoxy-D-manno-2-octulosonic acid and N-acetylneuraminic acid) and of various O-specific polysaccharides (from Aeromonas hydrophila and Aeromonas salmonicida) to the bifunctional spacer 1,6-hexanediamine, was achieved by reductive amination. The saccharide--1-(6-amino)-hexane alkyamines obtained were converted into the corresponding isothiocyanate derivatives and coupled to the free epsilon-amino group of lysine residues of the protein carrier bovine serum albumin. In similar manner, the aldehyde group introduced by selective periodate oxidation into the partially O-deacylated lipopolysaccharide of Vibrio anguillarum was conjugated to 1,6-hexanediamine, converted into the corresponding isothiocyanate and covalently attached to bovine serum albumin.

Structural elucidation of the O-specific polysaccharide of the phenol-phase soluble lipopolysaccharide of Vibrio anguillarum.

The structure of the O-specific polysaccharide of the phenol-soluble cellular lipopolysaccharide of Vibrio anguillarum has been investigated. The studies involved the use of methylation analysis, partial hydrolysis with 48% hydrogen fluoride, Smith degradation, oxidation with chromium trioxide, and comprehensive proton and carbon-13 nuclear magnetic resonance studies, in which one- and two-dimensional experiments were carried out. As a result of these studies it is proposed that the O-specific polysaccharide of Vibrio anguillarum is composed of a regular heteropolymer, i.e., a main chain of (1----4)-linked 3-acetamido-3,6-dideoxy-beta-L-glucose residues alternately substituted through O-2 with side chain residues of 2-acetamido-2,6-dideoxy-alpha-D-glucose, which seem to be substituted through either O-3 or O-4 with propionyl groups (R), as in the following structure. (Formula: see text)

Structure of the lipopolysaccharide core isolated from a human strain of Aeromonas hydrophila.

Lipopolysaccharide was isolated from the cell-walls of a human strain of Aeromonas hydrophila by the aqueous phenol method in 0.58% yield (based on dry weight of bacteria). The lipopolysaccharide consisted of SR-polysaccharide, core-oligosaccharide and lipid A; there was no O-specific polysaccharide. The core had the composition D-galactose, D-glucose, D-glycero-D-manno-heptose, L-glycero-D-manno-heptose and D-glucosamine in a molar ratio of 1:1:2:4:1. Glucosamine was linked to an L-glycero-D-manno-heptose residue by a bond which was resistant to hydrolysis. The D-glucosamine-(1----7)-LD-heptose disaccharide was isolated and identified by the mass spectrum of its methylated alditol and the heptose residue not observed under normal hydrolysis conditions was easily determined after deamination of the complete core. Methylation analysis, chemical degradation, periodate and chromium trioxide oxidations and nuclear magnetic resonance (13C and 1H NMR) spectroscopy were used to identify the structure of the core oligosaccharide as: (formula: see text)

Hydrolytic release, and identification by g.l.c.-m.s., of 3-deoxy-D-manno-2-octulosonic acid in the lipopolysaccharides isolated from bacteria of the Vibrionaceae.

The identification of the peracetylated methyl glycosides of 3-deoxy-D-manno-2-octulosonic acid (KDO) methyl esters was achieved by g.l.c.-m.s. These peracetylated methyl glycoside methyl esters were obtained from fully acetylated lipopolysaccharides and core oligosaccharides of representative strains of the Vibrionaceae family by the following sequence of mild reactions: acetolysis, methanolysis, and acetylation. KDO was shown to be present in all of the lipopolysaccharides (LPS), a result in direct contrast to the generally accepted view of the absence of this compound in LPS from this family of bacteria.

Chromium trioxide oxidation of acetylated 3-amino-3,6-dideoxy-L-hexosides.

Glycosides of the fully acetylated 3-amino-3,6-dideoxy-L-hexosides in the gluco, galacto, manno, and talo configurations have been treated with chromium trioxide in acetic acid. The alpha-L-methyl glycosides, which exist in the more stable chair conformation 1C4, with an axially oriented aglycon, are resistant to this oxidation, whereas the beta-L-linked glycoside having the gluco configuration (1C4 conformation) and the alpha-L-methyl glycoside having the talo configuration (4C1 conformation), both with equatorially oriented aglycons, are oxidized to the corresponding 5-adulosonates.