复合益生菌液态发酵黄芪的制备方法

Preparation Method of Compound Probiotic Liquid-state Fermented Astragalus

为研究复合益生菌液态发酵黄芪的制备方法,提高发酵液在临床应用中的作用效果,利用排列组合的方法,将枯草芽孢杆菌、酵母菌、乳酸菌按顺序接种至同一培养基,培养一定时间后观察各菌种的生长情况,确定菌种接种顺序;然后再用单因子试验法,以发酵液中菌体数量为指标,确定枯草芽孢杆菌的发酵时间,乳酸菌和酵母菌的接种时间、接种量以及发酵时间;最后,按优化的培养条件进行中试扩大试验,分别测定发酵液中各种菌体的数量。结果显示,利用多菌种对黄芪提取液进行混合发酵的制备方法能够实现,接种顺序为枯草芽孢杆菌、酵母菌、乳酸菌;最优发酵条件为:枯草芽孢杆菌以3. 0%的接种量,于37℃培养24 h后,以2. 0%的接种量接入酵母菌,于28℃培养48 h后,再以0. 5%的接种量接入乳酸菌,于37℃静置培养24 h;中试扩大试验所得发酵液中,枯草芽孢杆菌、酵母菌、乳酸菌含量分别为28. 52×10~8、5. 71×10~8、9. 34×10~8cfu/mL,终末pH值为4. 55。因此,利用复合益生菌液态发酵黄芪的制备方法具有较好的可行性。

The aim was to establish the stable preparation method for liquid-state fermentation of Astagulus by compound probiotic and improve the effect of fermentation liquor in clinical application.The inoculation order of Bacillus subtilis,yeast,and Lactobacillus was arranged with permutation and combination method.After cultivation for a definite time,the optimal inoculation order was confirmed by the growth situation of three strains.Furthermore,single factor test was used,with the number of probiotic in fermentation as the index,to measure the fermentation time of Bacillus subtilis,as well as the inoculation time,inoculation quantity and fermentation time of Lactobacillus and yeast.Finally,a pilot scale test,which referred to the optimized culture conditions,was applied to assay the quantity of probiotic in fermentation liquid.The results showed that compound probiotic liquid-state fermented Astragalus was feasible with the order as Bacillus subtilis,yeast and Lactobacillus.The optimum fermentation conditions were as follows:Culture of Bacillus subtilis at 37℃for 24 h with 3.0%incubating volume was cultured at 28℃for another 48 h with 2.0%incubating volume of yeast,followed by static incubation at 37℃for 24 h with 0.5%incubating volume of Lactobacillus.The contents of Bacillus subtilis,yeast and Lactobacillus in fermenting liquor from the pilot test were 28.52×10^8,5.71×10^8,9.34×10^8 cfu/mL,respectively.The fermenting liquor pH was 4.55.In conclusion,this study constructs a feasible preparation method of compound probiotic liquid-state fermented Astragalus.