【目的】探讨不同动物肠道优势需氧菌对黄豆苷原转化菌株转化能力的影响。【方法】有氧条件下,采用稀释涂布法分别从ICR小鼠、芦花鸡、长白猪和獭兔等4种健康动物肠道中分离优势需氧菌,将不同动物的优势需氧菌分别与不同类型黄豆苷原转化菌株进行厌氧混合培养,高效液相色谱检测培养液中黄豆苷原的转化情况。【结果】16S rRNA基因序列分析,结合形态学及相关理化特性分析表明,分离的22株优势需氧菌分属埃希氏菌属(10株)、变形菌属(5株)、肠球菌属(4株)、芽胞杆菌属(2株)和假单胞菌属(1株)五个属。混菌培养结果显示,兔源蜡样芽胞杆菌(R1)和铜绿假单胞菌(R5)与转化菌株混合并连续转接2-3次后,转化菌株的转化活性完全丧失,而其它用于混合培养的肠道优势需氧菌对转化菌株转化能力均无明显影响。菌株R1和R5分别与30只ICR小鼠肠道菌群在人工模拟肠道营养液中混合并连续5次转接培养后,约90%的ICR小鼠肠道菌群完全丧失将黄豆苷原转化为雌马酚的能力。【结论】不同动物肠道优势需氧菌对黄豆苷原转化菌株转化能力有不同影响,兔源蜡样芽胞杆菌R1和铜绿假单胞菌R5对转化菌株的转化能力有明显抑制作用。
[Objective] To investigate the influence of isolated predominant aerobic bacteria on daidzein biotransformation capacity by co-culture with daidzein biotransforming bacteria.[Methods] Predominant aerobic bacteria were isolated from diluted feces solutions of different healthy animals,including ICR mice,Luhua chicken,Landrace pigs and Rex rabbits.Daidzein biotransforming bacteria were anaerobically co-cultured with the isolated predominant aerobic bacteria and the cultural broth was extracted and detected by high performance liquid chromatography(HPLC).[Results] Twenty two predominant aerobic bacteria were isolated from the four different healthy animals mentioned above.Based on the analyses of 16S rRNA gene sequences,morphology study and relative biophysico-biochemical characteristics,all 22 isolates belong to the 5 genera,i.e.Escherichia(10),Proteus(5),Enterococcus(4),Bacillus(2) and Pseudomonas(1).Co-culture between predominant aerobic bacteria and daidzein biotransforming bacteria was carried out under anaerobic conditions.The results showed that the biotransformation capacity was totally lost when different daidzein biotransforming bacterium was co-cultured with either Bacillus cereus(R1) or Pseudomonas aerginosa(R5) and continuously inoculated for 2 or 3 passages.However,no obvious influence was observed when daidzein biotransforming bacteria were co-cultured with all the other isolated predominant aerobic bacteria except R1 and R5.In addition,when strain R1 and R5 was co-cultured with the intestinal microflora of the ICR mice anaerobically and continuously inoculated for 5 passages,about 90% of the co-cultures totally lost the activity to convert daidzein to equol effectively.[Conclusion] Different predominant aerobic bacteria showed different influence on daidzein biotransformation capacity after being co-cultured with different daidzein biotransforming bacteria.Among all the isolated predominant aerobic bacteria used for co-culture,both Bacillus cereus(R1) and Pseudomon