The Study Of Tartary Buckwheat Protein On Regulation Of Intestinal Flora

In this paper, tartary buckwheat as research material and the protein extracted by ammonium sulfate from tartary buckwheat as research object. The effect of tartary buckwheat protein on the growth and tolerate bile acid salts of probiotics (L.plantarum and L.acidophilus) were investigated by detecting the turbidity (OD600) of fermentation liquid in this paper through the pure culture in vitro. On this basis, through simulating the gastrointestinal tract digestion and colon fermentation in vitro, the number and structure of main intestinal microflora (Bifidobacterium, Lactobacillus, Enterococcus, Enterobacteriaceae, C.perfringens, Clostridium and Bacteroidetes), fermentation liquid pH and short chain fatty acids (acetic acid, propionic acid, butyric acid and lactic acid) content was measured by the method of conventional culture count, fluorescence quantitative PCR, PCR-TGGE combined with gas chromatography-mass spectrometry analysis. The major results were as follows:(1) The additional different concentrations of tartary buckwheat protein (1,3 and 5 mg/ml) increased the optical density value (OD600) of Lplantarum culture liquid by 12,26 and 35% respectively, and the optical density value (OD600) of L.acidophilus culture liquid by 15,27 and 31% respectively. Simultaneously, the results of Lplantarum and L.acidophilus tolerate to sodium cholate and sodium glycyl cholate increased significantly (p<0.05), and the OD600 values was 1.6,8.7 and 4.1,8.30 times of that of control, and those rusults reveals the tartary buckwheat protein had a certain role in promoting on the growth of Lactobacillus.(2) When cultured 32 h through simulating in vitro fermentation, traditional culture count results demonstrated tartary buckwheat protein can significantly promote the proliferation of Bifidobacteria and Lactobacillus and inhibit the growth of Enterococcus, Enterobacteriaceae and Clostridium (p<0.05). Such as the tartary buckwheat protein of high-dose group, increased the number of bifidobacteria, lactobacilli by 5.9,4.3 times respectively and decreased the number of Enterococcus, Escherichia coli and Clostridium by 65,72 and 78% respectively compared with the control group. The results of quantitative PCR were consistent with the results of traditional culture count. In addition, HD group increased the content of Clostridium content by one order of magnitude, and the inhibition rate of growth for Bacteroidetes was 85%.(3) The tartary buckwheat protein may also reduce the intestinal pH and significantly promote the metabolism of short chain fatty acids (p<0.05). HD group increased acetic acid, propionic acid, butyric acid, lactic acid by 8.9,33.8,27.3 and 25.6 times, respectively.(4) Tartary buckwheat protein could increase the diversity of the intestinal flora. With the concentration increase of buckwheat protein, their diversity also increased, in addition, its role in promoting significantly higher than the casein group (p<0.05).In summary, the tartary buckwheat protein can promote the growth of probiotics such as Bifidobacteria and reduce the proliferation of Enterococcus and other harmful bacteria, and also reveals the pathway of inhibition was related to tartary buckwheat protein can improve intestinal short chain fatty acids and reduce the pH of the intestinal.Thus, tartary buckwheat may act as a functional factor with probiotic function.