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Dextran without reactions.

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A W WILKINSON. 1956-09-22. Dextran without reactions.. https://doi.org/10.1016/s0140-6736(56)92294-2

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Structural and physicochemical characterisation of branched dextrans produced by an active α-(1→2) branching sucrase from Apilactobacillus kunkeei PDER37.

Recently, branching sucrases encoded in the genomes of certain Lactic Acid Bacteria (LAB) strains have become novel enzymes to obtain branched α-glucans. In this study an active α-(1 → 2) branching sucrase from Apilactobacillus kunkeei PDER37 was expressed, characterised and distinct branched dextrans was obtained with reactions under different sucrose: dextran ratio. Structural characterisation by 1H and 13C NMR analysis demonstrated the branching of the dextran with (1 → 2)-linked α-d-glucose units with no alteration in the final structure depending on sucrose: dextran ratio (D0) but this ratio was effective for the determination of the molecular weights of the branched dextrans (D1, D2 and D3). FTIR analysis further supported the dextran structures and suggested the higher accumulation of the α-Glc units in the branched dextrans. Thermal characterisation of the branched dextrans obtained by TGA and DSC analysis suggested the increased hygroscopicity of the branching units. Both SEM and AFM analysis demonstrated more porous chain like structures in the branched dextrans. This study provides valuable information on the role of active α-(1 → 2) branching sucrase (BS37) for the production of branched dextrans with potential increased physicochemical status applicable for food and other industries.

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Dextran molecular size and degree of branching as a function of sucrose concentration, pH, and temperature of reaction of Leuconostoc mesenteroides B-512FMCM dextransucrase.

Reactions of Leuconostoc mesenteroides B-512FMCM dextransucrase with increasing concentrations of sucrose, from 0.1 to 4.0 M, gave a decreasing amount of high-molecular weight dextran (HMWD) (>10(6) Da) with a concomitant increase in low-molecular weight dextran (LMWD) (<10(5) Da). At 0.1 M sucrose, pH 5.5, and 28 degrees C, 99.8% of the dextran had a MW>10(6) Da and at 4.0 M sucrose, 69.9% had a MW<10(5) Da and 30.1% had a MW>10(6) Da, giving a bimodal distribution. The degree of branching increased from 5% for 0.1 M sucrose to 16.6% for 4.0 M sucrose. The temperature had very little effect on the size of the dextran, which was >10(6) Da, but it had a significant effect on the degree of branching, which was 4.8% at 4 degrees C and increased to 14.7% at 45 degrees C. Both the molecular weight (MW) and the degree of branching were not significantly affected by different pH values between 4.5 and 6.0.

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Capillary electrophoresis of proteins in dextran-coated columns.

A simple coating technique by using uncross-linked dextran has been developed for fused-silica capillaries to be used in capillary electrophoresis of basic proteins. The capillaries were first silanized with a heterobifunctional silane (gamma-aminopropyltriethoxylsilane), which served as a coupling agent between the capillary inner wall and the polysaccharide coating. Dextran of high molecular mass (about 70 kDa) was activated with 1,1'-carbonyldiimidazole. Then the activated dextran was coupled to the primary amino groups that were anchored onto the inner wall of the silanized capillaries. The residual reactive groups on the dextran were further substituted by neutral functions in a coupling reaction with excess ethanolamine. By using dimethyl sulfoxide (DMSO) rather than aqueous buffer as the reaction medium, the extent of substitution was improved by minimizing the residual reactive groups at the surface. Since they are ionogenic, the electrosmotic flow in the system is relatively low. The chemically bound dextran coating showed good reproducibility and stability. In electrophoretic experiments basic proteins were separated with high efficiency by use of the dextran-coated fused-silica capillary columns. The main advantage of the method described here is that both polysaccharide activation and amine-coupling reactions were carried out under mild conditions at room temperature without catalysts. For this reason, the method is recommended to coat the inner wall of microfluidic separation channels which would not tolerate a harsh treatment.

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