本文選題：富鉻酵母 + 葡萄糖耐量因子　； 參考：《天然產(chǎn)物研究與開發(fā)》2016年11期

【摘要】：為探究葡萄糖耐量因子(GTF)分離純化工藝,簡(jiǎn)化純化步驟,以期得到GTF純品,本實(shí)驗(yàn)在前人氨水提取基礎(chǔ)上,采用有機(jī)溶劑沉淀除去雜蛋白,在有機(jī)溶劑的種類、溶劑的濃度、提取組分以及沉淀時(shí)間的確定等方面進(jìn)行了試驗(yàn),以有機(jī)鉻含量(μg)與總蛋白含量(mg)的比值為純化評(píng)價(jià)標(biāo)準(zhǔn),高相液相色譜對(duì)純化結(jié)果進(jìn)行驗(yàn)證,同時(shí)將純化前后的GTF作用于胰島素抵抗型Hep-G2肝癌細(xì)胞,測(cè)定細(xì)胞葡萄糖消耗量,檢測(cè)GTF純化品對(duì)細(xì)胞葡萄糖代謝的調(diào)節(jié)活性。最終確定用30%低溫乙醇沉淀20 min,10000×g離心取上清液分離純化GTF,此方法相較于氨水直接提取,有機(jī)鉻含量與總蛋白含量的比值提高了6倍多,GTF提取率和純度都得到了顯著提高,且純化后的GTF對(duì)胰島素抵抗型Hep-G2細(xì)胞葡萄糖代謝具有顯著調(diào)節(jié)作用(P0.01)。
[Abstract]:In order to explore the separation and purification process of glucose tolerance factor (GTF) and simplify the purification procedure, we used organic solvent to precipitate and remove the impurity protein on the basis of ammonia water extraction, and to simplify the purification process, and to obtain the pure GTF product. The extraction components and the determination of precipitation time were tested. The ratio of organic chromium content (渭 g) to total protein content (渭 g) was used as the standard of purification evaluation. The purification results were verified by high phase liquid chromatography. At the same time, the purified GTF was applied to insulin-resistant Hep-G2 hepatoma cells, the glucose consumption of the cells was measured, and the regulating activity of the purified GTF on glucose metabolism of the cells was measured. It was determined that GTFs were separated and purified by centrifugation with 30% low-temperature ethanol for 20 min ~ 10000 脳 g centrifugation. Compared with direct extraction with ammonia, the ratio of organic chromium content to total protein content was increased by more than 6 times and the extraction rate and purity of GTF were significantly improved. The purified GTF could significantly regulate glucose metabolism in insulin resistant Hep-G2 cells.
【作者單位】： 天津科技大學(xué)食品工程與生物技術(shù)學(xué)院;中國(guó)農(nóng)業(yè)科學(xué)院農(nóng)產(chǎn)品加工研究所;
【基金】：國(guó)家自然科學(xué)基金(31371763)
【分類號(hào)】：R914
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本文編號(hào)：1923807