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A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin

机译:半试验规模的分离纯化大豆(Glycine max)凝集素的方法

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Availability of gram quantities of purified soybean lectin (SBL) to scientists will foster discovery of novel biomedical applications of the lectin and provide the opportunity to investigate the antinutritional effects of SBL in soybean-consuming food animals and poultry. Therefore, a semi-pilot-scale procedure for isolating and purifying SBL was designed. Defatted soyflour was extracted overnight with 0.9% NaCl at 4 degreesC. The extract obtained was filtered (0.45 mum membrane) and subjected to affinity chromatography using a column containing N-acetyl-D-galactosamine resin that is specific for SBL. Bound SBL was eluted off the column with 0.14 M galactose solution. The eluent was ultrafiltered (30 kDa), and the resulting solution (SBL and water) was freeze-dried. Electrophoretic analysis and hemagglutination assay revealed that the freeze-dried SBL was similar to Sigma-grade SBL in purity and activity (35 and 33 HU/mg protein, respectively). The procedure yielded 141 mg of SBL/100 g of soyflour. [References: 57]
机译:科学家可获得的克量的纯化大豆凝集素(SBL)将促进发现该凝集素的新型生物医学应用,并为研究SBL在食用大豆的食用动物和家禽中的抗营养作用提供机会。因此,设计了用于分离和纯化SBL的半中试方法。脱脂的大豆粉在4℃下用0.9%NaCl提取过夜。将获得的提取物过滤(0.45μm膜),并使用含有对SBL具有特异性的N-乙酰基-D-半乳糖胺树脂的柱进行亲和色谱。用0.14M的半乳糖溶液从柱上洗脱结合的SBL。将洗脱液超滤(30 kDa),并将所得溶液(SBL和水)冷冻干燥。电泳分析和血凝分析表明,冻干的SBL的纯度和活性与Sigma级SBL相似(分别为35 HU / mg和33 HU / mg蛋白质)。该程序产生141 mg SBL / 100 g大豆粉。 [参考:57]

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