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以脱脂豆粉为原料,经pH7.6磷酸溶液抽提、65℃热变性、硫酸铵分步沉淀等提取技术制备粗提液,之后再经过DEAE-52离子交换、亲和层析和葡聚糖凝胶过滤等纯化技术研究大豆胰蛋白酶抑制因子的分离纯化方法。结果表明,从脱脂豆粉中分离纯化的大豆胰蛋白酶抑制因子的比活力高达4 600 U.mg-1,提纯倍数为73.85。纯化的大豆胰蛋白酶抑制因子经SDS-PAGE电泳分析,呈现2条谱带,分子量分别为21.92和20.04 kDa,这2种蛋白均为大豆胰蛋白酶抑制因子。该法操作简便,分离纯化效果好,对大豆胰蛋白酶抑制因子的研究与生产有重要的参考价值。
The defatted soybean powder was taken as raw material, extracted by pH7.6 phosphoric acid solution, heat denatured at 65 ℃ and fractionated by ammonium sulfate for extraction. The crude extract was then subjected to DEAE-52 ion exchange, affinity chromatography, Purification of soybean trypsin inhibitor using purification technology such as sugar gel filtration. The results showed that the specific activity of soybean trypsin inhibitor isolated and purified from defatted soybean flour was up to 4 600 U.mg-1 and the purification fold was 73.85. The purified soybean trypsin inhibitor showed two bands by SDS-PAGE electrophoresis, with molecular weights of 21.92 and 20.04 kDa, respectively, and the two proteins were soybean trypsin inhibitor. The method has the advantages of simple operation, good separation and purification effect, and important reference value for the research and production of soybean trypsin inhibitor.