Partial characterization of dextran-degrading enzyme obtained from blue cheese

Degradation of dextran beads was observed when the water-soluble fraction of a blue cheese extract was applied to the top of a Sephadex G-150 or G-200 column. This phenomenon suggests the presence of a specific enzyme that can hydrolyze dextran. After removal of casein components from the blue cheese fraction, ammonium sulfate treatment and gel filtration chromatography were performed to isolate the enzyme fraction. The enzymatic products were analyzed by thin-layer chromatography and gel filtration chromatography and identified as isomaltooligosaccharides. The isoelectric point of this enzyme fraction was approximately 4.9, as determined by isoelectric focusing using Rotofor, and the molecular weight of the fraction was 65 kDa, as estimated by sodium dodecyl sulfate (SDS)-PAGE. Optimum pH for enzymatic activity was 5.0 to 5.3. A partial N-terminal amino acid sequence of 20 residues was determined to be ATPDEWRSRSIYFMLTDRGA from an enzyme fraction further purified by ion-exchange chromatography and native PAGE. This sequence showed a maximum homology of 80% with alpha-amylase or Taka amylase that originated from various microorganisms.     

采用分级超滤法精制右旋糖酐

通过采用分级超滤的方法以不同的分子质量范围对右旋糖酐粗品进行分离纯化 ,并分析右旋糖酐在不同分子质量范围内的分布。结果表明 ,分级超滤在不添加有机溶剂的前提下能更有效地控制右旋糖酐产品的分子质量分布 ,能够取代传统的分级乙醇沉淀法。基于质量衡算 ,分级超滤能更准确地反映右旋糖酐粗品的分子质量分布     

利用Maillard反应制备大豆蛋白-葡聚糖共价复合物

研究了大豆酸沉蛋白和葡聚糖共价键合制备反应及其产物乳化性能的变化.在干热条件下,两种大分子通过Maillard反应进行共价键合并通过聚丙烯酰胺凝胶电泳验证了大分子复合物的存在. 结果表明产物具有比大豆酸沉蛋白更高的对油/水乳状液的乳化能力,复合物在pH 3.0和pH 10.0时均能保持较好的乳化活性,并且在高温、高盐条件下乳化活性变化很小.     

大分子拥挤环境下多糖对蛋白肽的修饰研究

采用胃蛋白酶适度水解大豆分离蛋白得到蛋白肽,在大分子拥挤环境下通过Maillard反应制备蛋白肽-葡聚糖共价复合物(Hydrolated Soy Protein Isolated-Dextran Conjugates,HDC),采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术证实大豆7S球蛋白和葡聚糖发生了共价结合,并通过乳化剪切机、荧光分光光度计、差示扫描量热仪和粒度分布仪等对其接枝度、溶解性、热稳定性、抗氧化性和乳化性能进行系统分析。结果表明:大分子拥挤环境下大豆水解蛋白肽-葡聚糖60℃反应4 h就可以形成较高接枝度的共价复合物,溶解性较大豆分离蛋白(Soybean protein isolated,SPI)和大豆分离蛋白-葡聚糖(Soybean Protein Isolated-Dextran Conjugates,SDC)有所提高,热稳定性好,HDC的自由基清除率明显高于大豆水解蛋白和它与葡聚糖的混合物,达到66.39%,且羟自由基清除率高于维生素C,HDC的乳液粒径最小并具有优越的乳化稳定性。     

右旋糖酐-阿司匹林偶联物的合成

以右旋糖酐和乙酰水杨酰氯为原料,合成了高分子偶联物右旋糖酐 阿司匹林,并用FTIR、1HNMR对其结构进行了表征。考察了缚酸剂、右旋糖酐重均相对分子质量、反应温度和物料比对酯化反应的影响。结果表明,三乙胺和吡啶均可作为该酯化反应的缚酸剂,而浓氢氧化钠水溶液则不适合;乙酰水杨酰基的接入率随右旋糖酐重均相对分子质量的增大而减小,但随乙酰水杨酰氯与右旋糖酐的质量比的增加而增大;当选用Mw=20 000的右旋糖酐、缚酸剂三乙胺,反应温度在 60℃左右、m(乙酰水杨酰氯) /m(右旋糖酐 ) =3. 06时,乙酰水杨酰基的接入率和其有效转化率分别为 9. 30%和 3 .07%。     

Newly designed polyacrylamide/dextran gels for electrophoresis protein separation: synthesis and characterization

Newly designed gels for electrophoresis protein separation were synthesized from acrylamide, N,N′‐methylenebis (acrylamide) and dextran mixtures. Radical polymerization was initiated by ammonium persulfate and N,N,N′,N′‐tetramethylethylenediamine. The time dependence of absorbance during polymerization was monitored using UV‐visible spectroscopy. The exothermic polymerization process exhibited a sharp rise of temperature reminiscent of the Trommsdorff effect. The swelling kinetics of the synthesized gels was examined in deionized water and buffer solutions. One of the challenges was to find an alternative to commercial products, sold as mixtures with no detailed chemical contents, commonly used in sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS‐PAGE) for protein separation. For this reason, a systematic comparison was made of the properties of one of the most commonly used commercial gels, Duracryl^? from Genomics Solution Inc., and those of the synthesized polyacrylamide/dextran gels. Copyright © 2011 Society of Chemical Industry     

乙醇沉淀法测定葡聚糖浓度方法探讨

研究采用乙醇沉淀法测定葡聚糖浓度实验方法,结果表明：乙醇与葡聚糖溶液体积比大于等于1∶1时,溶液产生较大浊度;反应时间在30 min以上时,溶液浊度相对稳定;但大量实验研究结果表明：葡聚糖溶液浓度与浊度之间线性关系不好,且溶液产生的浊度重复性很差。     

Partial or total silylation of dextran with hexamethyldisilazane

The silylation reaction of dextran with 1,1,1,3,3,3-hexamethyldisilazane (HMDS) in DMSO was studied as the first step of the synthesis of new amphiphilic polyester-grafted dextrans. According to the experimental conditions, i.e. dextran molar weight, medium temperature and reaction time, HMDS/OH ratio, addition of a catalyst and co-solvent, partially or totally silylated dextrans were recovered. The highest silylation yields were obtained with the lowest molecular weight dextrans. The increase in temperature medium and/or reaction time, the presence of catalyst or co-solvent favored the protection yield. Whatever the dextran used, complete silylation of the polysaccharide chain could be achieved by adequate selection of the experimental conditions. The thermal properties of resulting silylated polysaccharides were investigated by temperature modulated DSC. It was observed that T_g values of partially silylated dextran were maintained between 120 and 140 °C, independently of the dextran molecular weight. Interestingly, DMSO proved to behave as an efficient plasticizer of (partially) silylated dextrans. The partially silylated dextrans were efficiently used as multifunctional macroinitiators for the controlled ring-opening polymerization (ROP) of lactone. The ROP was then promoted from the remaining hydroxyl groups in the presence of tin or aluminium activator. After polymerization and ultimate deprotection of the silylated dextran backbone, amphiphilic polyester-grafted dextrans were readily recovered.     

Reproducible,biocompatible medical materials from biologically derived polysaccharides: Processing and Characterization

Natural polysaccharides like chitosan and dextran have garnered considerable interest in biomedical applications due to their biocompatibility, biodegradability, and nontoxicity. Nonetheless, the development of a reproducible class of medical devices from these materials is challenging and has had limited success. Chitosan and dextran are inherently variable and synthesis using these materials is prone to inconsistencies. In this study, we put forward a robust product development regimen that allows these natural materials to be developed into a reproducible class of biomaterials. First, an array of validated characterization methods (Proton Nuclear Magnetic Resonance, titrations, Ultraviolet spectroscopy, Size Exclusion Chromatography—Multi-Angle Light Scattering, Size Exclusion Chromatography—Refractive Index, and proprietary methods) were developed that allowed rigorous specifications to be set for unprocessed chitosan and dextran, chitosan and dextran intermediates, and chemically modified materials—acrylated chitosan (aCHN) and oxidized dextran (oDEX). Second, a robust and reproducible synthesis scheme involving various in-process controls was developed to chemically modify the unprocessed polysaccharides. Third, purification methods to remove byproducts and low-molecular-weight impurities for both aCHN and oDEX were developed. The study presents a viable strategy for converting variable, natural materials into a reproducible class of biomaterials that can be applied in various biomedical applications. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48454.