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以高铬酵母菌粉为研究对象,采用温和的纯化方法,通过氨水提取高铬酵母菌粉中的酵母菌葡萄糖耐量因子(glucose tolerance factor,GTF),联用SuperdexTM75和Sephadex G25凝胶过滤层析对提取物进行粗分离,然后用反相高效液相色谱对提取物进行再分离,从而对GTF进行高效纯化研究。将GTF纯化品作用于胰岛素抵抗型3T3-L1脂肪细胞,通过检测细胞葡萄糖消耗量,分析GTF纯化品对细胞葡萄糖代谢调节活性。研究表明,GTF纯化品可以有效提高胰岛素抵抗型3T3-L1脂肪细胞葡萄糖的消耗。可见,经过多步柱层析纯化后,可以得到具有葡萄糖代谢调节活性的GTF纯化品。
High-chromium yeast powder was used as the research object. The mild glucose purification method was used to extract glucose tolerance factor (GTF) of yeast from high-chromium yeast powder by supercritical ammonia water extraction. SuperdexTM75 and Sephadex G25 gel filtration chromatography Crude extract was separated, and the extract was separated by reversed-phase high-performance liquid chromatography to further purify GTF. The GTF purified product was applied to insulin-resistant 3T3-L1 adipocytes, and glucose metabolic activity of the purified GTF was assayed by measuring the glucose consumption of the cells. Studies have shown that GTF purified products can effectively increase the glucose consumption of insulin-resistant 3T3-L1 adipocytes. It can be seen that after purification by multi-step column chromatography, a GTF purified product with glucose metabolism-regulating activity can be obtained.