小麦麸皮中β-葡聚糖的分离纯化及组成研究
The isolation, purification and composition analysis of β-glucan from wheat bran
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摘要: 以小麦麸皮为原料,碱法提取β-葡聚糖。采用硫酸铵沉淀法、DEAE Sepharose CL-6B阴离子交换柱层析法、Sepharose CL-4B凝胶过滤层析法对β-葡聚糖的粗提物进行逐步纯化,得到组分A1和A2。并经紫外分光光度法、高效凝胶渗透色谱法鉴定其纯度,最终获得相对分子质量为4.87×105(纯度为98.57%)的β-葡聚糖A1组分、6.13×104左右(纯度为97.03%)的A2组分。可见,经过一系列的纯化实验,可以获得纯度较高的β-葡聚糖单一组分。采用特异性β-葡聚糖酶对经阴离子交换柱层析获得的β-葡聚糖组分A进行水解,经高效液相色谱法分析其寡糖的组成。结果表明,特异性酶解后的β-葡聚糖组分A主要由麦芽三糖和麦芽四糖组成,除了含有少量葡萄糖外,还含有麦芽糖、麦芽五糖。Abstract: With wheat bran as the raw material, β-glucan was extracted by alkaline extraction.The crude extraction of β-glucan was purified gradually by adding ( NH4) 2SO4and using DEAE Sepharose CL- 6B anion- exchange chromatography as well as Sepharose CL- 4B gel filtration chromatography, and components A1 and A2ofβ-glucan were obtained after purification. The purity of β- glucan was researched by UV scanning and high performance gel permeation chromatography analysis.The relative molecular weights of β-glucan components A1, A2 were respectively 4.87 × 105 ( purity was 98.57%) , 6.13 ×104 ( purity was 97.03%) .So homogeneous β-glucan components with high purity were obtained after purification. After β- glucanase enzymatic hydrolysis β- glucan component A from anion- exchange chromatography, the oligosaccharide composition was analysised by HPLC.The results showed that: the oligosaccharide constitutions were primarily maltotriose and maltotetraose. What's more, there were a small amount of glucose, maltose and maltopentaose.